LOCUS       sequence-430555-        7432 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV transfer plasmid to express stGtACR2-FusionRed in a 
            frt-dependent manner.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7432)
  AUTHORS   Kirchgessner MA, Franklin AD, Callaway EM
  TITLE     Distinct ""driving"" versus ""modulatory"" influences of different 
            visual corticothalamic pathways.
  JOURNAL   Curr Biol. 2021 Dec 6;31(23):5121-5137.e7. doi: 
            10.1016/j.cub.2021.09.025. Epub 2021 Oct 5.
  PUBMED    34614389
REFERENCE   2  (bases 1 to 7432)
  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"; doi: 
            "10.1016/j.cub.2021.09.025"; journalName: "Curr Biol"; date: 
            "2021-12-6- 6"; volume: "31"; issue: "23"; pages: "5121-5137.e7"
COMMENT     Sequence Label: pAAV-EF1a-fDIO-stGtACR2-FusionRed
FEATURES             Location/Qualifiers
     source          1..7432
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        231..1409
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          462..1400
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1357..1377
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    1440..1473
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     CDS             complement(1767..2462)
                     /codon_start=1
                     /product="monomeric red fluorescent protein optimized for 
                     use as a fusion tag (Shemiakina et al., 2012)"
                     /label=FusionRed
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMPMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     VVEGGPLPFAFDILATSFMYGSRTFIKHPPGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKVRGVNFPANGPVMQKKTLGWEASTETMYPADGGLEGACDMAL
                     KLVGGGHLICNLETTYRSKKPATNLKMPGVYNVDHRLERIKEADDETYVEQHEVAVARY
                     STGGAGDGGK"
     regulatory      3401..3410
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(3417..3450)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     misc_feature    3559..4147
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3612..3632)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4030..4041)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4179..4655
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4378..4397)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4695..4835
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4695..4824
                     /label=AAV2 ITR
     rep_origin      4910..5365
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4997..5016)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5207..5228
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5382..5401)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5501..5523
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5561..5579)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5647..5751
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5752..5820,5821..6612)
                     /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(5970..5989)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6783..7371
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7272..7291
                     /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 gcgtaagctt tgcaaagatg gataaagttt taaacagaga
      181 ggaatctttg cagctaatgg accttctagg tcttgaaagg agtgggaatt ggctccggtg
      241 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
      301 gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
      361 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
      421 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt
      481 cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct
      541 ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga
      601 ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct
      661 ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt
      721 ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc
      781 ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg
      841 acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac
      901 cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc
      961 cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag
     1021 cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct
     1081 cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg
     1141 tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga
     1201 gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc
     1261 acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg
     1321 aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa
     1381 gtttttttct tccatttcag gtgtcgtgag gtaccggatc ctctagagtc gactccggag
     1441 aagttcctat tctctagaaa gtataggaac ttcgcagaat ggtagctgga ttgtagctgc
     1501 tattagcaat atgaaacctc ttagaagttc ctattcttca aaaggtatag gaacttcggc
     1561 gcgccttact taaaccgcgt agcctctgga aaatcggtgg cgcatgagat aaaagagtca
     1621 atagaactca tggacctcat gtcgatcacg ccctctgttc ttgcaggaag aggtgccaca
     1681 ggggagggca ttgaggacat ttccagctcc tcagggggtt tactctgggg ggccatctct
     1741 ttggtgttca gaataggctg ggactgttta cctccatcac cagcgccacc agtagagtat
     1801 ctggccacag ccacctcgtg ctgctcgacg tatgtctcat cgtcggcctc cttgattctt
     1861 tccagtctgt ggtccacgtt gtagacgccg ggcatcttga ggttcgtagc gggtttcttg
     1921 gatctgtatg ttgtctcaag gttgcagatc aggtgtcctc ctcccacgag cttcagagcc
     1981 atgtcacatg cgccttccag tcctccgtca gcagggtaca tcgtctcggt ggaggcctcc
     2041 cagccgagtg ttttcttctg catcacaggg ccgttggctg ggaagttcac ccctctaacc
     2101 ttgacgttgt agatgaggca gccgtcctgg aggctggtgt cctgggtagc ggtcagcacg
     2161 cccccgtctt cgtatgtggt gactctctcc catgtgaagc cctcagggaa ggactgctta
     2221 aagaagtcgg ggatgcccgg agggtgcttg atgaaggttc tgctgccgta catgaagctg
     2281 gtagccagga tgtcgaaggc gaaggggaga gggccgccct cgacgacctt gattctcatg
     2341 gtctgggtgc cctcgtaggg cttgccttcg ccctcggatg tgcacttgaa gtggtggttg
     2401 ttcacggtgc cctccatgta cagcttcatg ggcatgttct ccttaatcag ctcgctcacc
     2461 atcaccacgt tgatgtcgat ctggtccagg gggatgtact cgccctcgct ggtgatcctg
     2521 ctcttggcgg cccttccaaa cattctgata ttgacttcgt cgttaggagg ggcattcttc
     2581 tcgtgattca ggccaaaagg atcgtcggct tcgggtttct cctgccctgg cagacatgcg
     2641 gggtcccact tcccttccag cactccccag gcagtgctcc acaggatcac tccatatgta
     2701 tttttgcaca ggccgtcagc cagcaggtac agaatggcgc tcacgttctc gtggatcacg
     2761 cagactccgg tgggtgaaat cagccacacg attgggaaca tagtccaagc aataaagaag
     2821 atcaggcgca gcatcttgat cctcaccacc accttctgtg ccagaggtgt cccgacagat
     2881 tcaaagccat acagccccac ttgaaagatt gtgaagacca cactgtactc gaacaccagg
     2941 caggtggccc caaagaaaaa gaacagccac ttcatatact ctgcgggaga aattgtagcg
     3001 gtgattccca tgaccactct aatgatggaa gcggcctgtg caatgggatt cagtgggata
     3061 cttttgtact tcaccagagc catgttagaa atctgtccca gcatgattgg acaggtgcac
     3121 agccaggagg ccattcggct ccaaggaaag aatctgccgt ccctcatctg cagggttccg
     3181 ttgccagtac tggccagtgc ataagtgata gattcgacaa agggcaggta cacggactcc
     3241 caagggacct tgggagccct gctaatcatg aagattgtct ggcagcctga gaccacggcc
     3301 cacaccagct gcagcagtga gacgaaggtg gagtcgattc tggacatatt gtagcactcg
     3361 gtgtgagttg atgcccattc cccatagacg acctgtgatg ccatggtggc gctagcgaag
     3421 ttcctatact ttctagagaa taggaacttc ttgccttaac ccagaaatta tcactgttat
     3481 tctttagaat ggtgcaaaga gaagttccta taccttttga agaataggaa cttcgaattc
     3541 gatatcaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     3601 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     3661 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     3721 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     3781 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     3841 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     3901 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg
     3961 tcctttcctt ggctgctcgc ctatgttgcc acctggattc tgcgcgggac gtccttctgc
     4021 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     4081 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     4141 tccccgcatc gataccgagc gctgctcgag agatctacgg gtggcatccc tgtgacccct
     4201 ccccagtgcc tctcctggcc ctggaagttg ccactccagt gcccaccagc cttgtcctaa
     4261 taaaattaag ttgcatcatt ttgtctgact aggtgtcctt ctataatatt atggggtgga
     4321 ggggggtggt atggagcaag gggcaagttg ggaagacaac ctgtagggcc tgcggggtct
     4381 attgggaacc aagctggagt gcagtggcac aatcttggct cactgcaatc tccgcctcct
     4441 gggttcaagc gattctcctg cctcagcctc ccgagttgtt gggattccag gcatgcatga
     4501 ccaggctcag ctaatttttg tttttttggt agagacgggg tttcaccata ttggccaggc
     4561 tggtctccaa ctcctaatct caggtgatct acccaccttg gcctcccaaa ttgctgggat
     4621 tacaggcgtg aaccactgct cccttccctg tccttctgat tttgtaggta accacgtgcg
     4681 gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg
     4741 ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc
     4801 ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct
     4861 tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt
     4921 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     4981 agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     5041 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     5101 cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga
     5161 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     5221 caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg
     5281 ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     5341 aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc
     5401 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     5461 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     5521 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt
     5581 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     5641 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     5701 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     5761 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     5821 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     5881 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     5941 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     6001 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     6061 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     6121 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     6181 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     6241 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     6301 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     6361 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     6421 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     6481 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     6541 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     6601 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     6661 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     6721 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     6781 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     6841 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     6901 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     6961 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     7021 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     7081 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     7141 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     7201 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     7261 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     7321 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     7381 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gt
//
