LOCUS       sequence-348374-        6744 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SOUL in Cre-positive cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6744)
  AUTHORS   Gong X, Mendoza-Halliday D, Ting JT, Kaiser T, Sun X, Bastos AM, 
            Wimmer RD, Guo B, Chen Q, Zhou Y, Pruner M, Wu CW, Park D, 
            Deisseroth K, Barak B, Boyden ES, Miller EK, Halassa MM, Fu Z, Bi G,
            Desimone R, Feng G
  TITLE     An Ultra-Sensitive Step-Function Opsin for Minimally Invasive 
            Optogenetic Stimulation in Mice and Macaques.
  JOURNAL   Neuron. 2020 Jul 8;107(1):197. doi: 10.1016/j.neuron.2020.06.018.
  PUBMED    32645306
REFERENCE   2  (bases 1 to 6744)
  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: "Neuron."; 
            date: "2020-07-8"; volume: "107(1):197. doi"; pages: " 
            10.1016/j.neuron.2020.06.018"
COMMENT     Sequence Label: pAAV-EF1a-FLEX-SOUL-P2A-tdTomato-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..6744
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        156..367
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     protein_bind    complement(395..428)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(479..512)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(541..1971)
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGIGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     CDS             complement(1972..2028)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             complement(2047..2973)
                     /codon_start=1
                     /product="H134R mutant of an N-terminal fragment of 
                     channelrhodopsin-2 from the alga Chlamydomonas reinhardtii 
                     (Nagel et al., 2005)"
                     /label=ChR2(H134R)
                     /note="blue light-activated cation channel"
                     /translation="MDYGGALSAVGRELLFVTNPVVVNGSVLVPEDQCYCAGWIESRGT
                     NGAQTASNVLQWLAAGFSILLLMFYAYQTWKSTCGWEEIYVCAIEMVKVILEFFFEFKN
                     PSMLYLATGHRVQWLRYAEWLLTSPVILIHLSNLTGLSNDYSRRTMGLLVSAIGCIVWG
                     ATSAMATGYVKVIFFCLGLCYGANTFFHAAKAYIEGYHTVPKGRCRQVVTGMAWLFFVS
                     WGMFPILFILGPEGFGVLSVYGSTVGHTIIDLMSKNCWGLLGHYLRVLIHEHILIHGDI
                     RKTTKLNIGGTEIEVETLVEDEAEAGAV"
     regulatory      2970..2979
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2992..3025
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    3076..3109
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3134..3722
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3187..3207)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3605..3616)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3756..3867
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3985..4125
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3985..4114
                     /label=AAV2 ITR
     rep_origin      4200..4655
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4287..4306)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4497..4518
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4672..4691)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4791..4813
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4851..4869)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4937..5041
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5042..5110,5111..5902)
                     /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(5260..5279)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6073..6661
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6562..6581
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(6723..6744,1..108)
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 ctcgctcact gaggccgccc gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg
       61 agcgagcgcg cagagaggga gtggccaact ccatcactag gggttcctgc ggccgcacgc
      121 gtaaggagtg ggaattggct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt
      181 ccccgagaag ttggggggag gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg
      241 ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga
      301 accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag
      361 aacacaggta ggatcctcta gagtcgactc cggaataact tcgtatagga tactttatac
      421 gaagttatgc agaatggtag ctggattgta gctgctatta gcaatatgaa acctcttaat
      481 aacttcgtat agcatacatt atacgaagtt atggcgcgcc gtcgactcgc gattcgcgcc
      541 ttacttgtac agctcgtcca tgccgtacag gaacaggtgg tggcggccct cggagcgctc
      601 gtactgttcc acgatggtgt agtcctcgtt gtgggaggtg atgtccagct tggtgtccac
      661 gtagtagtag ccgggcagtt gcacgggctt cttggccatg tagatggtct tgaactccac
      721 caggtagtgg ccgccgtcct tcagcttcag ggcctggtgg atctcgccct tcagcacgcc
      781 gtcgcggggg tacaggcgct cggtggaggc ctcccagccc atggtcttct tctgcattac
      841 ggggccgtcg ggggggaagt tggtgccgcg catcttcacc ttgtagatca gcgtgccgtc
      901 ctgcagggag gagtcctggg tcacggtcac cagaccgccg tcctcgaagt tcatcacgcg
      961 ctcccacttg aagccctcgg ggaaggacag cttcttgtaa tcggggatgt cggcggggtg
     1021 cttcacgtac gccttggagc cgtacatgaa ctggggggac aggatgtccc aggcgaaggg
     1081 cagggggccg cccttggtca ccttcagctt ggcggtctgg gtgccctcgt aggggcggcc
     1141 ctcgccctcg ccctcgatct cgaactcgtg gccgttcatg gagccctcca tgcgcacctt
     1201 gaagcgcatg aactctttga tgacggccat gttgttgtcc tcggaggagg cggtgccgga
     1261 gctgccgctg ccggtgctgc cgatgccatg ccccaggaac aggtggtggc ggccctcgga
     1321 gcgctcgtac tgttccacga tggtgtagtc ctcgttgtgg gaggtgatgt ccagcttggt
     1381 gtccacgtag tagtagccgg gcagttgcac gggcttcttg gccatgtaga tggtcttgaa
     1441 ctccaccagg tagtggccgc cgtccttcag cttcagggcc tggtggatct cgcccttcag
     1501 cacgccgtcg cgggggtaca ggcgctcggt ggaggcctcc cagcccatgg tcttcttctg
     1561 cattacgggg ccgtcggggg ggaagttggt gccgcgcatc ttcaccttgt agatcagcgt
     1621 gccgtcctgc agggaggagt cctgggtcac ggtcaccaga ccgccgtcct cgaagttcat
     1681 cacgcgctcc cacttgaagc cctcggggaa ggacagcttc ttgtaatcgg ggatgtcggc
     1741 ggggtgcttc acgtacgcct tggagccgta catgaactgg ggggacagga tgtcccaggc
     1801 gaagggcagg gggccgccct tggtcacctt cagcttggcg gtctgggtgc cctcgtaggg
     1861 gcggccctcg ccctcgccct cgatctcgaa ctcgtggccg ttcatggagc cctccatgcg
     1921 caccttgaag cgcatgaact ctttgatgac ctcctcgccc ttgctcacca ttggcccggg
     1981 attctcttcg acatcccctg cttgtttcaa cagggagaag ttagtggctc cgcttccgtt
     2041 aactggcacg gctccggcct cggcttcgtc ttcgacgaga gtctcgacct cgatctccgt
     2101 tccgccgatg ttcagtttgg tggttttgcg gatatctccg tgaatcaata tgtgctcgtg
     2161 gatcaggacg cgcaggtagt gtcccaacaa cccccaacaa tttttactca tcagatcaat
     2221 aatcgtgtga cctacggtgg agccatagac gctcaggacg ccaaaacctt cgggccccaa
     2281 aatgaagaga attgggaaca taccccagct cacgaaaaac agccatgcca tgccggtcac
     2341 gacctggcgg caccgaccct ttggcacagt atgataaccc tcgatatatg ctttggcggc
     2401 gtgaaaaaat gtgttcgcgc catagcacaa tccaagacaa aagaagatga ctttaacata
     2461 gccggttgcc atggcgctgg tagcccccca cacgatacac ccgatggctg agacaaggag
     2521 tcccatggtt ctcctgctgt agtcgttgct caggccggtg aggttgctca ggtggataag
     2581 gatgacagga gaagtgagca gccactctgc atagcgcagc cactgcaccc ggtgtcctgt
     2641 ggcaaggtag agcatagagg gattcttaaa ctcaaaaaag aactcgagaa tcaccttaac
     2701 catttcaatg gcgcacacat agatctcctc ccagccgcat gtagatttcc aggtttggta
     2761 ggcatagaac atcagcagca aaatgctgaa tcctgctgca agccactgca ggacatttga
     2821 cgcggtctga gcgccgttcg tgccgcgaga ttcaatccat ccggcacagt aacattgatc
     2881 ctcagggacc aggacggacc cgttcaccac cacaggatta gtaacgaaca aaagttcgcg
     2941 tccgacggca gacaaagcgc cgccatagtc catggtggcg gccgcgctag cataacttcg
     3001 tataaagtat cctatacgaa gttatttgcc ttaacccaga aattatcact gttattcttt
     3061 agaatggtgc aaagaataac ttcgtataat gtatgctata cgaagttatg aattcgatat
     3121 caagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     3181 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     3241 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     3301 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     3361 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     3421 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     3481 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt
     3541 tccttggctg ctcgcctatg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     3601 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     3661 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     3721 gcatcgatac cgagcgctgc tcgagagatc ttcgactgtg ccttctagtt gccagccatc
     3781 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     3841 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     3901 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgagat
     3961 ctcacgtgcg gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc
     4021 tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg
     4081 cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt
     4141 attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta
     4201 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     4261 tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     4321 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     4381 tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc
     4441 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     4501 actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata
     4561 agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     4621 cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg
     4681 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     4741 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     4801 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
     4861 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     4921 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     4981 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     5041 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     5101 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     5161 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     5221 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     5281 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     5341 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     5401 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     5461 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     5521 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     5581 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     5641 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     5701 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     5761 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     5821 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     5881 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     5941 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     6001 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     6061 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     6121 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     6181 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     6241 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     6301 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     6361 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     6421 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     6481 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     6541 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     6601 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     6661 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     6721 gtcctgcagg cagctgcgcg ctcg
//
