LOCUS       sequence-360819-        7884 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector to express nuclear localized E2-Crimson from PAG 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7884)
  AUTHORS   de Solis CA, Hosek MP, Holehonnur R, Ho A, Banerjee A, Luong JA, 
            Jones LE, Chaturvedi D, Ploski JE
  TITLE     Adeno-associated viral serotypes differentially transduce inhibitory
            neurons within the rat amygdala.
  JOURNAL   Brain Res. 2017 Oct 1;1672:148-162. doi: 
            10.1016/j.brainres.2017.07.023. Epub 2017 Jul 29.
  PUBMED    28764932
REFERENCE   2  (bases 1 to 7884)
  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: "Brain 
            Res."; date: "2017-10-1"; volume: "1672:148-162. doi"; pages: " 
            10.1016/j.brainres.2017.07.023. Epub 2017 Jul 29"
COMMENT     Sequence Label: pAAV-PAG-E2-Crimson-3XNLS-pA
FEATURES             Location/Qualifiers
     source          1..7884
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          2639..3114
                     /label=beta-globin intron
                     /note="internally truncated intron from human beta-globin"
     regulatory      3211..3220
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3217..3891
                     /codon_start=1
                     /product="far-red noncytotoxic tetrameric variant of DsRed 
                     fluorescent protein (Strack et al., 2009)"
                     /label=E2-Crimson
                     /note="mammalian codon-optimized"
                     /translation="MDSTENVIKPFMRFKVHMEGSVNGHEFEIEGVGEGKPYEGTQTAK
                     LQVTKGGPLPFAWDILSPQFFYGSKAYIKHPADIPDYLKQSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGTLIYHVKFIGVNFPSDGPVMQKKTLGWEPSTERNYPRDGVLKGENHM
                     ALKLKGGGHYLCEFKSIYMAKKPVKLPGYHYVDYKLDITSHNEDYTVVEQYERAEARHH
                     LFQ"
     primer_bind     3615..3634
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             3904..3924
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3928..3948
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3952..3972
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        complement(4014..4032)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4015..4032)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4052..4069)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4058..4169
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4313..4442
                     /label=AAV2 ITR
     rep_origin      4813..5326
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4900..4919)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5110..5131
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5808..5827)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5927..5949
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5987..6005)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6073..6177
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6178..6246,6247..7038)
                     /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(6396..6415)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7209..7797
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7698..7717
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
       61 gagcgcagcg agtcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct
      181 agaggatcct tcgaaaagct agcacgcgtc ctctacccac tgagccatgt cctaagttcc
      241 taattgttaa atatactaga acgagtttaa atctttaaca tcagaagtca tttccttgaa
      301 caatggttga gaaagtagga aaaactggca tgaaaacgtt gactggatta gtgaatagtc
      361 tggatgaggt tgagaaaaag gaaaacagag gcagtaaaaa actgattagt tgtttttaag
      421 aaactgagca acactaaggt ttggaagcca cccatcagtg aaagaaaaag ttaggtatta
      481 ttaagggtta cacatgctat ctctctccca caatgtgatg tttttcttaa aaacattata
      541 actttgattt actaataata tatatttttg agaagttaac cttaagccat aatcaaagcc
      601 acccaagtaa tttacaattc tgtttttttt ttctacaagg tcacttccct cacatttttc
      661 tatctaaaga gcaggttgaa aaaacaactt gaacacggca atcctgtatg taatacaaca
      721 tgtaggcaca gttagattct ttggttacta atccagttca gcatttacag attcattgga
      781 ttactaaatc tggtgaatta tgcccttccc atctcataaa tatcctgcta accttggtga
      841 atctattcat ttaagccatt ttgtttcaga agatggttta aaaacatttt tattctgaag
      901 cggaatgtgt gtacaggctc aggaactata tgttgcacag atctaagaag aaatggatgg
      961 agtgatgctg aaacagtttt gagacagggt agtcagatgc ttcattttaa aacaggtttt
     1021 aaaatagaaa agggatctgc agtatatttt gctttcaagc caaccagtac taatgaaata
     1081 gtcactgttc aaaaatctat tggagatact gcaaagaaac tgggggagat gtggtctgag
     1141 caatctgcca aagataaaca accgtatgag cagaaagcag ctaaactaga ggagaagtat
     1201 gaaaaggata ttgctgcata ccgtgccggg gcaaaagtga agtaggaaag aagggtcctg
     1261 gtaggccaac aggctcaaag aagaatgaac cagaagatga ggaagaggag gaggaggaag
     1321 atgatgaaaa tgaagaggag gaagatgagg ctgaagagta agtatctgtc ctaaagtgtg
     1381 gagtatatgt gctcaggcaa ttattttgct aagaatgtga aattcaagtg cagctcaaca
     1441 ttagcttcag tataaaaact gtacagattt ttgtatagct gattagactc tttgtagaga
     1501 aaatactttt taaaaagagt ttgtagcttt ttcaggggct acaacgtaca gttagatgta
     1561 aagcttttga tgttgaatgt ttctaaatat ttaatggttt ctttaatttc ttatgatagc
     1621 aaaaaaaaaa cttcatagga atttgtatta ccagtaaaag aatttttttt aggatgttgc
     1681 atttttgttt ttttttaatt tgtaataaaa taatgtatat aaaaaatttg gagaaatcgt
     1741 gtagatgata gaagagatcc acccttactc tggcttttct cttttcttcg ttgttgctac
     1801 gctcaaccgt tctaaatgtc ttaagttgca tagctgaaag gattttgctc attttttcca
     1861 aaacatttcg tttctgggca gtagccaacc atggaaagtt ttaagcacat ttgggagagg
     1921 gatgaagctc cttagggcgg ccttggcagg accgacgcct tcagaagggc agacacattc
     1981 ccctagccga gggaatgagc aattggcttg gataaggcga tcggccaccg ccggcctccc
     2041 tccagccctg ggaggcccgg actggcgcac tccagcttcc tcaatagctc gaggacgtgc
     2101 ttccctggag cgtacctgcc tggtgaatag ccagactccg gagccgcggg tcctcgccca
     2161 gcgtggcctc aggtggcccg aagcgagggg aagagcggct ggcggggctg acacacgtca
     2221 gtcgcccttc tccccgccct ccaggacccg cgggactgcg cacgggcgcg gagacgaatt
     2281 tgactgattc gctcgccaat cagaagagag gtcggcaggc tgagccaatg ggagggcgcg
     2341 cagggggcag accgtcggcg tcctcccgcc ctcggttggt gagagcggta tgtctccagc
     2401 cctcccctgc gcgtccgtct cagtgcggag ctcgcggcgg ccggaggaac ttaggctggt
     2461 ggcaggctcg gcggcggcgg cggcgatagc ggcggaggag gagcaggggc cacaccccac
     2521 cgcggacttt tttcttctca gacctcgtcc cgccgtgcgg gacaccgttc tgggagaaga
     2581 gcggacgccc acgccccgag gatctggcgc gccgagctcg gatcctgaga acttcagggt
     2641 gagtctatgg gacccttgat gttttctttc cccttctttt ctatggttaa gttcatgtca
     2701 taggaagggg agaagtaaca gggtacacat attgaccaaa tcagggtaat tttgcatttg
     2761 taattttaaa aaatgctttc ttcttttaat atactttttt gtttatctta tttctaatac
     2821 tttccctaat ctctttcttt cagggcaata atgatacaat gtatcatgcc tctttgcacc
     2881 attctaaaga ataacagtga taatttctgg gttaaggcaa tagcaatatt tctgcatata
     2941 aatatttctg catataaatt gtaactgatg taagaggttt catattgcta atagcagcta
     3001 caatccagct accattctgc ttttatttta tggttgggat aaggctggat tattctgagt
     3061 ccaagctagg cccttttgct aatcatgttc atacctctta tcttcctccc acagctcctg
     3121 ggcaacgtgc tggtctgtgt gctggcccat cactttggca aagcacgtga attcgaagat
     3181 ctggccgcct cggcctctag aactagtgcc gccaccatgg atagcactga gaacgtcatc
     3241 aagcccttca tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca cgagttcgag
     3301 atcgagggcg tgggcgaggg caagccctac gagggcaccc agaccgccaa gctgcaagtg
     3361 accaagggcg gccccctgcc cttcgcctgg gacatcctgt ccccccagtt cttctacggc
     3421 tccaaggcgt acatcaagca ccccgccgac atccccgact acctcaagca gtccttcccc
     3481 gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc
     3541 caggactcct ccctgcagga cggcaccctc atctaccacg tgaagttcat cggcgtgaac
     3601 ttcccctccg acggccccgt aatgcagaag aagactctgg gctgggagcc ctccactgag
     3661 cgcaactacc cccgcgacgg cgtgctgaag ggcgagaacc acatggcgct gaagctgaag
     3721 ggcggcggcc actacctgtg tgagttcaag tccatctaca tggccaagaa gcccgtgaag
     3781 ctgcccggct accactacgt ggactacaag ctcgacatca cctcccacaa cgaggactac
     3841 accgtggtgg agcagtacga gcgcgccgag gcccgccacc acctgttcca gtctcgagct
     3901 gatccaaaaa agaagagaaa ggtagatcca aaaaagaaga gaaaggtaga tccaaaaaag
     3961 aagagaaagg taggatccac cggatctaga taacgtacgg gaaacggccc tattctatag
     4021 tgtcacctaa atgctagagc tcgctgatca gcctcgactg tgccttctag ttgccagcca
     4081 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     4141 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     4201 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag cgcggccgct
     4261 ctagagcatg gctacgtaga taagtagcat ggcgggttaa tcattaacta caaggaaccc
     4321 ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga
     4381 ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc
     4441 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     4501 aatggcgaat ggcgattccg ttgcaatggc tggcggtaat attgttctgg atattaccag
     4561 caaggccgat agtttgagtt cttctactca ggcaagtgat gttattacta atcaaagaag
     4621 tattgcgaca acggttaatt tgcgtgatgg acagactctt ttactcggtg gcctcactga
     4681 ttataaaaac acttctcagg attctggcgt accgttcctg tctaaaatcc ctttaatcgg
     4741 cctcctgttt agctcccgct ctgattctaa cgaggaaagc acgttatacg tgctcgtcaa
     4801 agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     4861 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     4921 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     4981 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     5041 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     5101 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     5161 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     5221 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttaa atatttgctt
     5281 atacaatctt cctgtttttg gggcttttct gattatcaac cggggtacat atgattgaca
     5341 tgctagtttt acgattaccg ttcatcgatt ctcttgtttg ctccagactc tcaggcaatg
     5401 acctgatagc ctttgtagag acctctcaaa aatagctacc ctctccggca tgaatttatc
     5461 agctagaacg gttgaatatc atattgatgg tgatttgact gtctccggcc tttctcaccc
     5521 gtttgaatct ttacctacac attactcagg cattgcattt aaaatatatg agggttctaa
     5581 aaatttttat ccttgcgttg aaataaaggc ttctcccgca aaagtattac agggtcataa
     5641 tgtttttggt acaaccgatt tagctttatg ctctgaggct ttattgctta attttgctaa
     5701 ttctttgcct tgcctgtatg atttattgga tgttggaatc gcctgatgcg gtattttctc
     5761 cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct
     5821 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     5881 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     5941 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     6001 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     6061 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     6121 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     6181 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     6241 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     6301 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     6361 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     6421 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     6481 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     6541 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     6601 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     6661 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     6721 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     6781 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     6841 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     6901 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     6961 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     7021 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     7081 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     7141 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     7201 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     7261 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     7321 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     7381 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     7441 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     7501 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     7561 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     7621 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     7681 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     7741 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     7801 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     7861 tttcctgcgt tatcccctga ttct
//
