LOCUS       sequence-434073-        5800 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  NLS-Gal4(1-65)-iLight2-VP16 construc expression in mammalian cells. 
            pAAV vector..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5800)
  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 5800)
  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-iLight2-VP16
FEATURES             Location/Qualifiers
     source          1..5800
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(18..38)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     repeat_region   155..284
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(346..934)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(426..445)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1105..1896,1897..1965))
                     /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     1728..1747
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1966..2070)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2138..2156
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2194..2216)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2316..2335
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2352..2807)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2489..2510)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2701..2720
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2882..3022
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2893..3022
                     /label=AAV2 ITR
     primer_bind     complement(3132..3151)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3333..3353
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3424..3657)
                     /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"
     promoter        complement(join(5789..5800,1..88))
                     /label=CMVd1 promoter
                     /note="truncated human cytomegalovirus (CMV) immediate 
                     early promoter that yields slightly reduced expression 
                     relative to the full-length promoter"
ORIGIN
        1 atagacctcc caccgtacac gcctaccgcc catttgcgtc aatggggcgg agttgttacg
       61 acattttgga aagtcccgtt gattttggca ggaaagaaca tgtgagcaaa aggaattctc
      121 agctagccca caagtgtgca gcatcttcgg ccgcaggaac ccctagtgat ggagttggcc
      181 actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc
      241 ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggacatgt gagcaaaagg
      301 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
      361 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
      421 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
      481 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
      541 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
      601 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
      661 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
      721 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
      781 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
      841 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
      901 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
      961 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     1021 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     1081 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     1141 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     1201 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     1261 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     1321 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     1381 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     1441 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     1501 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     1561 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     1621 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     1681 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     1741 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     1801 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     1861 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     1921 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     1981 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     2041 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     2101 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     2161 tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     2221 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     2281 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag
     2341 agtgcaccat aaaattgtaa acgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     2401 aatcagctca ttttttaacc aatagaccga aatcggcaaa atcccttata aatcaaaaga
     2461 atagcccgag atagagttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     2521 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     2581 accatcaccc aaatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     2641 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     2701 agggaagaaa gcgaaaggag cgggcgctaa ggcgctggca agtgtagcgg tcacgctgcg
     2761 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtact atggttgctt
     2821 tgacgtatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc
     2881 ccctgcaggc agctgcgcgc tcgctcgctc actgaggccg cccgggcaaa gcccgggcgt
     2941 cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc
     3001 aactccatca ctaggggttc ctgcggccgc tttaaaaaac ctcccacatc tccccctgaa
     3061 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     3121 ttacaaataa agcaatagca tcacaaattt cacaaataaa caccacggaa ttgtcagtgc
     3181 ccaacagccg agcccctgtc cagcagcggg caaggcaggc ggcgatgagt tccgccgtgg
     3241 caagaactaa ccaggattta tacaaggagg agaaaatgaa agccatacgg gaagcaatag
     3301 catgatacaa aggcattaaa gcagcgtatc cacatagcgt aaaaggagca acatagttaa
     3361 gaataccagt caatctttca caaattttgt aatccagagg ttgattatct cgagcctaga
     3421 ttacccaccg tactcgtcaa ttccaagggc atcggtaaac atctgctcaa actcgaagtc
     3481 ggccatatcc agagcgccgt agggggcgga gtcgtggggg gtaaatcccg gaccagggga
     3541 atccccgtcc cccaacatgt ccagatcgaa atcgtctagc gcgtcggcat gcgccatcgc
     3601 cacgtcctcg ccgtctaagt ggagctcgtc ccccaggctg acatcggtcg ggggggccgt
     3661 cgacagtctg cgcgtgtgtc ccgcggggag aaaggacagg cgcggagccg ccagccccgc
     3721 ctcttcgggg gcgtcgtcgt ccgggagatc gagcaggccc tcgatggtag acccgtaatt
     3781 gtttttcgta cgcgcgcggc tgtacgcaga tcttcctcct gatccaccac ctccagaacc
     3841 acctcctcct gatccaccac ctccagaacc tcctccacct gatccaccac ccccgctggt
     3901 accgccgcca gaggtcagat cgtcgaagga ggccagcttc tccagattct cgttggcatc
     3961 ggccagctgg tcattcagca ggttcagctg catgctatcg gccacgatca gcaggtcccg
     4021 ggcgatgtcc cgggctgcat acagctgtgc ggttctccaa ggctgggact tgccgctcac
     4081 ctcgtcctgc caggcctcaa aagacttccg tgggcccacc atggtgcctg tagaggtctc
     4141 cacggacagc ttctcaggct tgcctgccca tgtgcgcacc tcgttctgtg ccaccctgaa
     4201 cagcagcagg tagctaaaca gatcggcgtc agacttcagt gggatggcca gcagtcctgc
     4261 gaaccggctg ccatctggca gcagtccagg tgcgtcctcc ttcagcatgt gggagtgcca
     4321 aaacaggctc ttcttgccat tctcctccag ccactccacc agcttgttga ttgtggtctg
     4381 atcaggggtc tcgccatatg tggtcagctc ctcgcctctc aggtagccca cgccatcaca
     4441 ccggaacagc ttgcaccagt cggcggcgtg ctcgatcacg atctcgtggg agctgtgctt
     4501 atcgtcggct gtggtagaca gctgctcccg ggctgcctgc actgtcacca tgtaccgctc
     4561 cacattcctg gagtggatca gccacaggcg ctctgctgca aactccacgg tgcgcaccag
     4621 caggtgcctg attctccggc cgatggccct aggctttgtg tggtgacagg ccacgatgcc
     4681 ccacagcttg tcctcattga agatgccgat ggaggtgctg gcagacacgc caaagttgcg
     4741 cagatactgc atgtgcagtg gagacacggc cctcagcacg ccgcaggaca gattcacggc
     4801 ctctgtgttc tgtggcttgt ggatcatgtg cagaggctgg ggctcggcat tcacatcggg
     4861 gatgatgcgg attgggttga tgctgtacat ggccctggcc tgtgcaggga tgtcagaggc
     4921 tgggaagtgg tggttcagca tggaggtaaa cagctgtctc acgctctcgg cgatcactct
     4981 gccattccac tcgggatcga actgatagat catcactctg tcgtagccgc tgatctcctg
     5041 gatcagccgt gtcagggtat tgatcagatc gttgtgggtc ttacagtttc tcagcttctg
     5101 gaaggcccgc tgaaactgga tctcgaacca attggatgtc tgcacgtggt atctctccac
     5161 ctcgatgata tagtagttct cgtggcgcca gcccagaaag ctcagcctgt cggtctccac
     5221 ccagataggc tcgttctcag agatggcgga tgtcaggccg ctgatcagct gctcgggcag
     5281 cagagaattg atgttggcct gcttgaaatc ggacagctcg tggatggtat tgtcggccac
     5341 gctgaagtac tcggcgctgt tggcagaggc gtacacgatc tgctgtgtgt ccttctccac
     5401 gatcagcatg gcgccaaatg gctggatggc attggggatg tggattggct cctgatcgca
     5461 ggcggcgatc agcttgctga tatcgtcaga gcccaggtcg gcggcgaatt ccccaccagc
     5521 actagtctgt tccagtcttt ctagccttga ttccacttct gtcagatgtg ccctagtcag
     5581 cggagacctt ttggttttgg gagagtagcg acactcccag ttgttcttca gacacttggc
     5641 gcacttcggt ttttctttgg agcacttgag ctttttaagt cggcaaatat cgcatgcttg
     5701 ttcgatagaa gacagtagct ttccaccacc tccggacttc ctcttcttct tgcgcgccat
     5761 gctagcggat ctgacggttc actaaaccag ctctgcttat
//
