LOCUS       sequence-438062-        7143 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV-STARR-seq validation control vector with TFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7143)
  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 7143)
  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_TFP_WPRE_polyA
FEATURES             Location/Qualifiers
     source          1..7143
                     /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..3224
                     /codon_start=1
                     /product="monomeric teal (cyan) fluorescent protein (Ai et 
                     al., 2006)"
                     /label=mTFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEETTMGVIKPDMKIKLKMEGNVNGHAFVIEGEGEGKPYDG
                     TNTINLEVKEGAPLPFSYDILTTAFAYGNRAFTKYPDDIPNYFKQSFPEGYSWERTMTF
                     EDKGIVKVKSDISMEEDSFIYEIHLKGENFPPNGPVMQKKTTGWDASTERMYVRDGVLK
                     GDVKHKLLLEGGGHHRVDFKTIYRAKKAVKLPDYHFVDHRIEILNHDKDYNKVTVYESA
                     VARNSTDGMDELYK"
     misc_feature    3268..3856
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3321..3341)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3739..3750)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3859..3875)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3860..3876)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3890..4366
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4089..4108)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4406..4546
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4406..4535
                     /label=AAV2 ITR
     rep_origin      4621..5076
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4708..4727)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4918..4939
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5093..5112)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5212..5234
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5272..5290)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5358..5462
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5463..5531,5532..6323)
                     /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(5681..5700)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6494..7082
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6983..7002
                     /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 gtcatggtga
     2521 gcaagggcga ggagaccaca atgggcgtaa tcaagcccga catgaagatc aagctgaaga
     2581 tggagggcaa cgtgaatggc cacgccttcg tgatcgaggg cgagggcgag ggcaagccct
     2641 acgacggcac caacaccatc aacctggagg tgaaggaggg agcccccctg cccttctcct
     2701 acgacattct gaccaccgcg ttcgcctacg gcaacagggc cttcaccaag taccccgacg
     2761 acatccccaa ctacttcaag cagtccttcc ccgagggcta ctcttgggag cgcaccatga
     2821 ccttcgagga caagggcatc gtgaaggtga agtccgacat ctccatggag gaggactcct
     2881 tcatctacga gatacacctc aagggcgaga acttcccccc caacggcccc gtgatgcaga
     2941 agaagaccac cggctgggac gcctccaccg agaggatgta cgtgcgcgac ggcgtgctga
     3001 agggcgacgt caagcacaag ctgctgctgg agggcggcgg ccaccaccgc gttgacttca
     3061 agaccatcta cagggccaag aaggcggtga agctgcccga ctatcacttt gtggaccacc
     3121 gcatcgagat cctgaaccac gacaaggact acaacaaggt gaccgtttac gagagcgccg
     3181 tggcccgcaa ctccaccgac ggcatggacg agctgtacaa gtaaggatcc tctagagtcg
     3241 acctgcagaa gcttcaagct tatcgataat caacctctgg attacaaaat ttgtgaaaga
     3301 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     3361 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     3421 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     3481 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     3541 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     3601 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     3661 aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
     3721 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     3781 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
     3841 tgggccgcct ccccgcatcg ataccgtcga cctcgagacc tagatctacg ggtggcatcc
     3901 ctgtgacccc tccccagtgc ctctcctggc cctggaagtt gccactccag tgcccaccag
     3961 ccttgtccta ataaaattaa gttgcatcat tttgtctgac taggtgtcct tctataatat
     4021 tatggggtgg aggggggtgg tatggagcaa ggggcaagtt gggaagacaa cctgtagggc
     4081 ctgcggggtc tattgggaac caagctggag tgcagtggca caatcttggc tcactgcaat
     4141 ctccgcctcc tgggttcaag cgattctcct gcctcagcct cccgagttgt tgggattcca
     4201 ggcatgcatg accaggctca gctaattttt gtttttttgg tagagacggg gtttcaccat
     4261 attggccagg ctggtctcca actcctaatc tcaggtgatc tacccacctt ggcctcccaa
     4321 attgctggga ttacaggcgt gaaccactgc tcccttccct gtccttctga ttttgtaggt
     4381 aaccacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct
     4441 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
     4501 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg
     4561 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt
     4621 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     4681 ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     4741 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     4801 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc
     4861 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     4921 gactcttgtt ccaaactgga acaacactca actctatctc gggctattct tttgatttat
     4981 aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta
     5041 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct
     5101 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     5161 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     5221 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     5281 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     5341 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     5401 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     5461 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     5521 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     5581 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     5641 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     5701 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     5761 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     5821 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     5881 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     5941 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     6001 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     6061 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     6121 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     6181 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     6241 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     6301 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     6361 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     6421 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     6481 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     6541 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     6601 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     6661 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     6721 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     6781 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     6841 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     6901 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     6961 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     7021 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     7081 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     7141 tgt
//
