LOCUS       sequence-365690-        5422 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector packaging AkaLuciferase under the TRE promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5422)
  AUTHORS   Kasatkina LA, Ma C, Matlashov ME, Vu T, Li M, Kaberniuk AA, Yao J, 
            Verkhusha VV
  TITLE     Optogenetic manipulation and photoacoustic imaging using a 
            near-infrared transgenic mouse model.
  JOURNAL   Nat Commun. 2022 May 19;13(1):2813. doi: 10.1038/s41467-022-30547-6.
  PUBMED    35589810
REFERENCE   2  (bases 1 to 5422)
  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: "Nat 
            Commun."; date: "2022-05-19"; volume: "13(1):2813. doi"; pages: " 
            10.1038/s41467-022-30547-6"
COMMENT     Sequence Label: pAAV-TRE-Tight2-AkaLuc
FEATURES             Location/Qualifiers
     source          1..5422
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     44..68
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      103..112
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1823..1843)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   2203..2343
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2203..2332
                     /label=AAV2 ITR
     rep_origin      2418..2873
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2505..2524)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2715..2736
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2890..2909)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3009..3031
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3069..3087)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3155..3259
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3260..3328,3329..4120)
                     /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(3478..3497)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4291..4879
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4780..4799
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4941..5070
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     protein_bind    5104..5125
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5140..5145,5146..5163,5164..5170)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     promoter        join(5179..5422,1..71)
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    5187..5205
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5223..5241
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5258..5276
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5294..5312
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5330..5348
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5365..5383
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5401..5419
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
ORIGIN
        1 tatgtcgagg taggcgtgta cggtgggagg cctatataag cagagctcgt ttagtgaacc
       61 gtcagatcgc ctggagaatt cgagctcggt acccggggat ccgccaccat ggaagatgcc
      121 aaaaacatta agaagggccc agcgccgttc tacccactcg aagacgggac cgccggcgag
      181 cagctgcaca aagccatgaa gcgctacgcc ctggtgcccg gcgccatcgc ctttaccgac
      241 gcacatattc aggtggacgt tacctacgcc gagtacttcg agatgagcgt tcggctggca
      301 gaagctatga ggcgctatgg gctgaataca aaccatcgga tcgtggtgtg cagcgagaat
      361 agctcgcagt tcttcatgcc cgtgttgggt gccctgttca tcggtgtggc tgtggcccca
      421 gctaacgaca tctacaacga gcgcgagctg ctgaacagca tgggcatcag ccagcccacc
      481 gtcgtattcg tgagcaagaa agggctgcga aaggtcctca acgtgcaaaa gaagctaccg
      541 atcatacgaa agatcatcat catggatagc aagaccgact accagggctt ccaaagcatg
      601 tacaccttcg tgacttccca tttgccaccc agcttcaacg agtacgactt cgtgcccgag
      661 agcttcgacc gggacaaaac catcgccctg atcatgaaca gtagtggtag tacaggatta
      721 cccaagggcg tagccctacc gcaccgcacc gcttgtgtcc gattcagtca tgcccgcgac
      781 cccatcttcg gctaccagaa catccccgac accgctatcc tcagcgtggt gccatttcac
      841 cacggcttcg gcatgttcac cacgctgggc tacttgatct gcggctttcg ggtcgtgctc
      901 atgtaccgct tcgaggagga gctattcttg cgcagcttgc aagactataa gattcaatct
      961 gccctgctgg tgcccacact atttagctgc ctcgctaaga gcactctcat cgacaagtac
     1021 gacctaagca gcttgcgcga gatcgccagc ggcggggcgc cgctcagcaa ggaggtaggt
     1081 gaggccgtgg ccaaacgctt ccgcctacca ggcatccgcc agggctatgg cctgacagaa
     1141 acaaccaacg ccgtcatgat cacccccgag ggggaccgta agcctggctc agtaggcaag
     1201 gtggtgccct tcttcgaggc taaggtggta gacttggtca ccggtaagac actgggtgtg
     1261 aaccagcgcg gtgagctgtg cgtccgtggc cccatgatca tgagcggcta cgttaacaac
     1321 cccgaggcta cgaacgctct catcgacaag gacggctggc tgcacagcgg cgacatcgcc
     1381 tactgggacg aggacgagca cttcttcatc gtggaccggc tgaagagcct gatcaaatac
     1441 aagggctacc aggtagcccc agccgaactg gagggcatcc tgctgcaaca cccctacatc
     1501 ttcgacgccg gagtcgccgg cctgcccgac gacgatgccg gcgagctgcc cgccgcagtc
     1561 gtcgtgttgg aacacggtaa aaccatgacc gagaaagaga tcgtggacta tgtggccagc
     1621 caggttacaa ccgccaagaa gctgcgcggt ggtgttgtgt ttgtggatga agtccctaga
     1681 ggatcgaccg gcaagttaga cgcccgcaag atccgcgaga ttctcactaa ggccaagaag
     1741 gacggcaaga tcgccgtgta actcgagata atcaacctct ggattacaaa atttgtgaaa
     1801 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     1861 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     1921 cctggttagt tcttgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     1981 gggctcggct gttgggcact gacaattccg tggtgtttat ttgtgaaatt tgtgatgcta
     2041 ttgctttatt tgtaaccatc tagctttatt tgtgaaattt gtgatgctat tgctttattt
     2101 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     2161 caggttcagg gggagatgtg ggaggttttt taaagcggcc gcaggaaccc ctagtgatgg
     2221 agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg
     2281 cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc
     2341 aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac
     2401 gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     2461 tacgcgcagc gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt
     2521 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     2581 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga
     2641 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     2701 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg
     2761 ctattctttt gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct
     2821 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg
     2881 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
     2941 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     3001 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     3061 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     3121 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     3181 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     3241 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     3301 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     3361 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     3421 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     3481 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     3541 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     3601 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     3661 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     3721 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     3781 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     3841 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     3901 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     3961 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     4021 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     4081 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     4141 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     4201 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     4261 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     4321 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     4381 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     4441 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     4501 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     4561 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     4621 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     4681 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     4741 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     4801 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     4861 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     4921 ctggcctttt gctcacatgt cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc
     4981 ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg
     5041 gagtggccaa ctccatcact aggggttcct gcggccgcag atgctgcaca cttgtgggct
     5101 agctaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
     5161 tcgtatgttg tgtggctcga gtttactccc tatcagtgat agagaacgta tgtcgagttt
     5221 actccctatc agtgatagag aacgatgtcg agtttactcc ctatcagtga tagagaacgt
     5281 atgtcgagtt tactccctat cagtgataga gaacgtatgt cgagtttact ccctatcagt
     5341 gatagagaac gtatgtcgag tttatcccta tcagtgatag agaacgtatg tcgagtttac
     5401 tccctatcag tgatagagaa cg
//
