LOCUS       sequence-438063-        7145 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV-STARR-seq validation vector with GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7145)
  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 7145)
  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: pssAAV_Seq17_CMV_bglobin_3NLS_GFP_WPRE_polyA
FEATURES             Location/Qualifiers
     source          1..7145
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        1331..1634
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1635..1838
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1789..1809
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1835..1859
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          1987..2365
                     /label=chimeric intron
                     /note="fusion between human cytomegalovirus intron A and 
                     human beta-globin intron 2"
     CDS             2445..2465
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2466..2486
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2493..2513
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(2514..2516,2517..2519,2520..3233)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2514..2516,2517..2519,2520..3230)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2559..2580)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2820..2839)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3167..3188
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3270..3858
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3323..3343)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3741..3752)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3861..3877)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3862..3878)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3892..4368
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4091..4110)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4408..4548
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4408..4537
                     /label=AAV2 ITR
     rep_origin      4623..5078
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4710..4729)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4920..4941
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5095..5114)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5214..5236
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5274..5292)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5360..5464
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5465..5533,5534..6325)
                     /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(5683..5702)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6496..7084
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6985..7004
                     /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 gcgtcttttc ttttttctaa tttattttaa tttattcact
      181 ttacatccca ctcactgccc cctcccactc acaccctccc acaatccttt ccccatcccc
      241 catcccccat ccccttttct tctgagcagg tgggggtccc atgggtattc cctaaccctg
      301 ccacttcaag tcttcctctc ccactgaggc gaggcaaggc agcccagcta gaacatatcc
      361 cacagacagg caacagcttt tgggatagcg ctcattctgg ttgtttagaa cccatatgaa
      421 gaccaagctg cacatctgct acattcgtaa aaggaagcct aggtccagcc tgtgtatgtt
      481 cattggttgg tagttcagtc tctgagaact ccaagggtcc aggttagttg attctattgg
      541 tcttcctgtg gagttcctat ccccttcggg gcctgcaatc tttcttccta ttcttcttta
      601 agagtctcca agctccatcc accgattgat tgtgagtgtc tgtatctgcc tgagtcagct
      661 gctgggtgga gcctctcaga gaacaacatg ctcctgtctt caagcataac agagtatcat
      721 taataatgta agggattggg cttgcccata ggatggatct caggacggtt attggttggc
      781 cattccgtca gtctctgctc cctgccccgt gcctacactt cgtgtagaca ggataaattt
      841 tgggttgaaa gttttgtggc tgggttggtg tctgtatcac tgcactgggg tacaggagat
      901 agcctcttca ggttccatat ccacaatgtt gtgaatcaca gtttcacagt taatgtcatc
      961 cccattgatt cttgctgccc ccttttcttc aaagttctct cagccttgac gggaggagtg
     1021 tgagacagac attacatcta gggctgagta ttctgtagtc tcttattctc tgcaccatgg
     1081 ccagttgtgg ttctctgcgt tacatgctat ctgcttcaaa aaagaagctt ccttgatggg
     1141 gcttgtttga tgcactaacc catgcaccat gtccatttag cagaaataat agtagattgg
     1201 ctctgctctc ttgacaaggg gtcatggtcc gtgcaatggc ctctgaactg accctcatta
     1261 tctacgcgtc tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
     1321 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
     1381 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
     1441 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     1501 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
     1561 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     1621 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
     1681 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
     1741 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
     1801 gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcg
     1861 cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc
     1921 tccgcggatt cgaatcccgg ccgggaacgg tgcattggaa cgcggattcc ccgtgccaag
     1981 agtgacgtaa gtaccgccta tagagtctat aggcccacaa aaaatgcttt cttcttttaa
     2041 tatacttttt tgtttatctt atttctaata ctttccctaa tctctttctt tcagggcaat
     2101 aatgatacaa tgtatcatgc ctctttgcac cattctaaag aataacagtg ataatttctg
     2161 ggttaaggca atagcaatat ttctgcatat aaatatttct gcatataaat tgtaactgat
     2221 gtaagaggtt tcatattgct aatagcagct acaatccagc taccattctg cttttatttt
     2281 atggttggga taaggctgga ttattctgag tccaagctag gcccttttgc taatcatgtt
     2341 catacctctt atcttcctcc cacagctcct gggcaacgtg ctggtctgtg tgctggccca
     2401 tcactttggc aaagaattgg gattcgaaca tcgataccat ggacccaaaa aagaagcgga
     2461 aggtcccaaa aaagaagcgg aaggtcgaat tcccaaaaaa gaagcggaag gtcatggtga
     2521 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
     2581 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
     2641 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
     2701 ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg
     2761 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
     2821 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
     2881 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
     2941 agtacaacta caacagccac aacgtctata tcatggccga caagcagaag aacggcatca
     3001 aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact
     3061 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
     3121 gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
     3181 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag taaggatcct
     3241 ctagagtcga cctgcagaag cttatcgata atcaacctct ggattacaaa atttgtgaaa
     3301 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     3361 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     3421 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     3481 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     3541 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     3601 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     3661 ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     3721 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     3781 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     3841 tttgggccgc ctccccgcat cgataccgtc gacctcgaga cctagatcta cgggtggcat
     3901 ccctgtgacc cctccccagt gcctctcctg gccctggaag ttgccactcc agtgcccacc
     3961 agccttgtcc taataaaatt aagttgcatc attttgtctg actaggtgtc cttctataat
     4021 attatggggt ggaggggggt ggtatggagc aaggggcaag ttgggaagac aacctgtagg
     4081 gcctgcgggg tctattggga accaagctgg agtgcagtgg cacaatcttg gctcactgca
     4141 atctccgcct cctgggttca agcgattctc ctgcctcagc ctcccgagtt gttgggattc
     4201 caggcatgca tgaccaggct cagctaattt ttgttttttt ggtagagacg gggtttcacc
     4261 atattggcca ggctggtctc caactcctaa tctcaggtga tctacccacc ttggcctccc
     4321 aaattgctgg gattacaggc gtgaaccact gctcccttcc ctgtccttct gattttgtag
     4381 gtaaccacgt gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct
     4441 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct
     4501 ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc
     4561 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata
     4621 gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     4681 cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     4741 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     4801 tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg
     4861 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     4921 tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt
     4981 ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt
     5041 taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat
     5101 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     5161 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     5221 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
     5281 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     5341 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     5401 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     5461 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     5521 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     5581 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     5641 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     5701 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     5761 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     5821 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     5881 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     5941 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     6001 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     6061 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     6121 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     6181 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     6241 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     6301 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     6361 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     6421 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     6481 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     6541 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     6601 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     6661 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     6721 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     6781 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     6841 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     6901 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     6961 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     7021 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     7081 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     7141 catgt
//
