LOCUS       sequence-434077-        5836 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  NLS-Gal4(1-65)-iLight-VP16 construc expression in mammalian cells. 
            pAAV vector..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5836)
  AUTHORS   Baloban M, Kasatkina LA, Verkhusha VV
  TITLE     iLight2: A near-infrared optogenetic tool for gene transcription 
            with low background activation.
  JOURNAL   Protein Sci. 2024 May;33(5):e4993. doi: 10.1002/pro.4993.
  PUBMED    38647395
REFERENCE   2  (bases 1 to 5836)
  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: "Protein 
            Sci."; date: "2024-05"; volume: "33(5):e4993. doi"; pages: " 
            10.1002/pro.4993"
COMMENT     Sequence Label: pAAV-CMVd1-NLS-GAL4-iLight-VP16
FEATURES             Location/Qualifiers
     source          1..5836
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     97..118
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(272..291)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     391..413
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(451..469)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        537..641
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(642..710,711..1502)
                     /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(860..879)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1673..2261
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2162..2181
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2323..2452
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        2519..2618
                     /label=CMVd1 promoter
                     /note="truncated human cytomegalovirus (CMV) immediate 
                     early promoter that yields slightly reduced expression 
                     relative to the full-length promoter"
     primer_bind     2569..2589
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             4786..5019
                     /codon_start=1
                     /gene="UL48"
                     /product="transcriptional activation domain of herpes 
                     simplex virus protein VP16 (Triezenberg et al., 1988; 
                     Cousens et al., 1989)"
                     /label=VP16 AD
                     /translation="APPTDVSLGDELHLDGEDVAMAHADALDDFDLDMLGDGDSPGPGF
                     TPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     primer_bind     complement(5090..5110)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     primer_bind     5292..5311
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   5421..5561
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5421..5550
                     /label=AAV2 ITR
     rep_origin      join(5636..5836,1..255)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5723..5742)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt
       61 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
      121 caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg
      181 tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
      241 taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
      301 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
      361 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
      421 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
      481 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
      541 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
      601 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
      661 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
      721 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
      781 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
      841 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
      901 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
      961 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     1021 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     1081 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     1141 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     1201 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     1261 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     1321 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     1381 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     1441 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     1501 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     1561 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     1621 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     1681 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     1741 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     1801 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     1861 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     1921 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     1981 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     2041 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     2101 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     2161 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     2221 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     2281 ttttacggtt cctggccttt tgctggcctt ttgctcacat gtcctgcagg cagctgcgcg
     2341 ctcgctcgct cactgaggcc gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg
     2401 agcgagcgag cgcgcagaga gggagtggcc aactccatca ctaggggttc ctgcggccga
     2461 agatgctgca cacttgtggg ctagctgaga attccttttg ctcacatgtt ctttcctgcc
     2521 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
     2581 gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc
     2641 gctagcatgg cgcgcaagaa gaagaggaag tccggaggtg gtggaaagct actgtcttct
     2701 atcgaacaag catgcgatat ttgccgactt aaaaagctca agtgctccaa agaaaaaccg
     2761 aagtgcgcca agtgtctgaa gaacaactgg gagtgtcgct actctcccaa aaccaaaagg
     2821 tctccgctga ctagggcaca tctgacagaa gtggaatcaa ggctagaaag actggaacag
     2881 actagtgctg gtgggtccgc cggcgggagt gctggcggta gtgccggcgg tgaattcgcc
     2941 gccgacctgg gctctgacga tatcagcaag ctgatcgccg cctgcgatca ggagccaatc
     3001 cacatcccca atgccatcca gccatttggc gccatgctga tcgtggagaa ggacacacag
     3061 cagatcgtgt acgcctctgc caacagcgcc gagtacttca gcgtggccga caataccatc
     3121 cacgagctgt ccgatttcaa gcaggccaac atcaattctc tgctgcccga gcagctgatc
     3181 agcggcctga catccgccat ctctgagaac gagcctatct gggtggagac cgacaggctg
     3241 agctttctgg gctggcgcca cgagaactac tatatcatcg aggtggagag ataccacgtg
     3301 cagacatcca attggttcga gatccagttt cagcgggcct tccagaagct gagaaactgt
     3361 aagacccaca acgatctgat caataccctg acacggctga tccaggagat cagcggctac
     3421 gacagagtga tgatctatca gttcgatccc gagtggaatg gcagagtgat cgccgagagc
     3481 gtgagacagc tgtttacctc catgctgaac caccacttcc cagcctctga catccctgca
     3541 caggccaggg ccatgtacag catcaaccca atccgcatca tccccgatgt gaatgccgag
     3601 ccccagcctc tgcacatgat ccacaagcca cagaacacag aggccgtgaa tctgtcctgc
     3661 ggcgtgctga gggccgtgtc tccactgcac atgcagtatc tgcgcaactt tggcgtgtct
     3721 gccagcacct ccatcggcat cttcaatgag gacaagctgt ggggcatcgt ggcctgtcac
     3781 cacacaaagc ctagggccat cggccggaga atcaggcacc tgctggtgcg caccgtggag
     3841 tttgcagcag agcgcctgtg gctgatccac tccaggaatg tggagcggta catggtgaca
     3901 gtgcaggcag cccgggagca gctgtctacc acagccgacg ataagcacag ctcccacgag
     3961 atcgtgatcg agcacgccgc cgactggtgc aagctgttcc ggtgtgatgg cgtgggctac
     4021 ctgagaggcg aggagctgac cacatatggc gagacccctg atcagaccac aatcaacaag
     4081 ctggtggagt ggctggagga gaatggcaag aagagcctgt tttggcactc ccacatgctg
     4141 aaggaggacg cacctggact gctgccagat ggcagccggt tcgcaggact gctggccatc
     4201 ccactgaagt ctgacgccga tctgtttagc tacctgctgc tgttcagggt ggcacagaac
     4261 gaggtgcgca catgggcagg caagcctgag aagctgtccg tggagacctc tacaggcacc
     4321 atggtgggcc cacggaagtc ttttgaggcc tggcaggacg aggtgagcgg caagtcccag
     4381 ccttggagaa ccgcacagct gtatgcagcc cgggacatcg cccgggacct gctgatcgtg
     4441 gccgatagca tgcagctgaa cctgctgaat gaccagctgg ccgatgccaa cgagaatctg
     4501 gagaagctgg cctccttcga cgatctgacc tctggcggcg gtaccagcgg gggtggtgga
     4561 tcaggtggag gaggttctgg aggtggtgga tcaggaggag gtggttctgg aggtggtgga
     4621 tcaggaggaa gatctgcgta cagccgcgcg cgtacgaaaa acaattacgg gtctaccatc
     4681 gagggcctgc tcgatctccc ggacgacgac gcccccgaag aggcggggct ggcggctccg
     4741 cgcctgtcct ttctccccgc gggacacacg cgcagactgt cgacggcccc cccgaccgat
     4801 gtcagcctgg gggacgagct ccacttagac ggcgaggacg tggcgatggc gcatgccgac
     4861 gcgctagacg atttcgatct ggacatgttg ggggacgggg attcccctgg tccgggattt
     4921 accccccacg actccgcccc ctacggcgct ctggatatgg ccgacttcga gtttgagcag
     4981 atgtttaccg atgcccttgg aattgacgag tacggtgggt aatctaggct cgagataatc
     5041 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     5101 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     5161 ctttcatttt ctcctccttg tataaatcct ggttagttct tgccacggcg gaactcatcg
     5221 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     5281 tgtttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     5341 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gagatgtggg
     5401 aggtttttta aagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg
     5461 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg
     5521 ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt
     5581 tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg
     5641 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     5701 cttgccagcg ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     5761 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     5821 ttacggcacc tcgacc
//
