LOCUS       sequence-438065-        7139 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV-STARR-seq validation vector with RFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7139)
  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 7139)
  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_RFP_WPRE_polyA
FEATURES             Location/Qualifiers
     source          1..7139
                     /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             2514..3227
                     /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    3264..3852
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3317..3337)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3735..3746)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3855..3871)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3856..3872)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3886..4362
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4085..4104)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4402..4542
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4402..4531
                     /label=AAV2 ITR
     rep_origin      4617..5072
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4704..4723)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4914..4935
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5089..5108)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5208..5230
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5268..5286)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5354..5458
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5459..5527,5528..6319)
                     /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(5677..5696)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6490..7078
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6979..6998
                     /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 cggaatggcc aaccaataac cgtcctgaga tccatcctat
      661 gggcaagccc aatcccttac attattaatg atactctgtt atgcttgaag acaggagcat
      721 gttgttctct gagaggctcc acccagcagc tgactcaggc agatacagac actcacaatc
      781 aatcggtgga tggagcttgg agactcttaa agaagaatag gaagaaagat tgcaggcccc
      841 gaaggggata ggaactccac aggaagacca atagaatcaa ctaacctgga cccttggagt
      901 tctcagagac tgaactacca accaatgaac atacacaggc tggacctagg cttcctttta
      961 cgaatgtagc agatgtgcag cttggtcttc atatgggttc taaacaacca gaatgagcgc
     1021 tatcccaaaa gctgttgcct gtctgtggga tatgttctag ctgggctgcc ttgcctcgcc
     1081 tcagtgggag aggaagactt gaagtggcag ggttagggaa tacccatggg acccccacct
     1141 gctcagaaga aaaggggatg ggggatgggg gatggggaaa ggattgtggg agggtgtgag
     1201 tgggaggggg cagtgagtgg gatgtaaagt gaataaatta aaataaatta gaaaaaagaa
     1261 aagacgcgtc 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 gtcatggtgt
     2521 ctaagggcga agagctgatt aaggagaaca tgcacatgaa gctgtacatg gagggcaccg
     2581 tgaacaacca ccacttcaag tgcacatccg agggcgaagg caagccctac gagggcaccc
     2641 agaccatgag aatcaaggtg gtcgagggcg gccctctccc cttcgccttc gacatcctgg
     2701 ctaccagctt catgtacggc agcagaacct tcatcaacca cacccagggc atccccgact
     2761 tctttaagca gtccttccct gagggcttca catgggagag agtcaccaca tacgaagacg
     2821 ggggcgtgct gaccgctacc caggacacca gcctccagga cggctgcctc atctacaacg
     2881 tcaagatcag aggggtgaac ttcccatcca acggccctgt gatgcagaag aaaacactcg
     2941 gctgggaggc caacaccgag atgctgtacc ccgctgacgg cggcctggaa ggcagaagcg
     3001 acatggccct gaagctcgtg ggcgggggcc acctgatctg caacttcaag accacataca
     3061 gatccaagaa acccgctaag aacctcaaga tgcccggcgt ctactatgtg gaccacagac
     3121 tggaaagaat caaggaggcc gacaaagaga cctacgtcga gcagcacgag gtggctgtgg
     3181 ccagatactg cgacctccct agcaaactgg ggcacaaact taattgagga tcctctagag
     3241 tcgacctgca gaagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga
     3301 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     3361 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     3421 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     3481 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     3541 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     3601 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat
     3661 catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
     3721 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     3781 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     3841 ccgcctcccc gcatcgatac cgtcgacctc gagacctaga tctacgggtg gcatccctgt
     3901 gacccctccc cagtgcctct cctggccctg gaagttgcca ctccagtgcc caccagcctt
     3961 gtcctaataa aattaagttg catcattttg tctgactagg tgtccttcta taatattatg
     4021 gggtggaggg gggtggtatg gagcaagggg caagttggga agacaacctg tagggcctgc
     4081 ggggtctatt gggaaccaag ctggagtgca gtggcacaat cttggctcac tgcaatctcc
     4141 gcctcctggg ttcaagcgat tctcctgcct cagcctcccg agttgttggg attccaggca
     4201 tgcatgacca ggctcagcta atttttgttt ttttggtaga gacggggttt caccatattg
     4261 gccaggctgg tctccaactc ctaatctcag gtgatctacc caccttggcc tcccaaattg
     4321 ctgggattac aggcgtgaac cactgctccc ttccctgtcc ttctgatttt gtaggtaacc
     4381 acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc
     4441 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc
     4501 gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt
     4561 ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc
     4621 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     4681 acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     4741 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     4801 tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc
     4861 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     4921 cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg atttataagg
     4981 gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     5041 gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc
     5101 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
     5161 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     5221 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg
     5281 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     5341 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     5401 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     5461 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     5521 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     5581 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     5641 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     5701 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     5761 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     5821 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     5881 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     5941 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     6001 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     6061 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     6121 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     6181 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     6241 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     6301 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     6361 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     6421 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     6481 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     6541 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     6601 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     6661 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     6721 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     6781 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     6841 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     6901 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     6961 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     7021 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     7081 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgt
//
