LOCUS       sequence-353739-        6994 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of the avian TVA receptor in a cre-dependent manner.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6994)
  TITLE     Ben-Simon lab CVS-N2c plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6994)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pAAV-DIO-CAG-TVA-P2A-dTomato
FEATURES             Location/Qualifiers
     source          1..6994
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   7..136
                     /label=AAV2 ITR
     primer_bind     1159..1181
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1243..1262
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    complement(1313..1346)
                     /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(1397..1430)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1448..2149)
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLYGMDELYK"
     primer_bind     complement(1531..1554)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(1729..1748)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1998..2016
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     CDS             complement(2150..2206)
                     /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"
     regulatory      2662..2671
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2678..2711
                     /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    2762..2795
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2820..3408
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2873..2893)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3291..3302)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3435..3452)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3441..3665
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        complement(3875..4204)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3889..4024
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4189..4209
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(4450..5038)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4530..4549)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5209..6000,6001..6069))
                     /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     5832..5851
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6070..6174)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6242..6260
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6298..6320)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6420..6439
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6653..6672
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        6962..6980
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     6962..6979
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 ttctagctgc gcgctcgctc gctcactgag gccgcccggg caaagcccgg gcgtcgggcg
       61 acctttggtc gcccggcctc agtgagcgag cgagcgcgca gagagggagt ggccaactcc
      121 atcactaggg gttccttgta gttaatgatt aacccgccat gctacttatc tacgtagcca
      181 tgctctaggt accattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      241 ctttccattg acgtcaatgg gtggagtatt trssswamwc yrycactwgg ggttccttgt
      301 agttaatgat taacccgcca tgctacttat ctacgtagcc atgctctagg taccattgac
      361 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
      421 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
      481 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
      541 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
      601 ggtcgaggtg agccccacgt tctgcttcac tctccccatc tcccccccct ccccaccccc
      661 aattttgtat ttatttattt tttaattatt ttgtgcagcg atgggggcgg gggggggggg
      721 ggggcgcgcg ccrggggggg gggggggggg gggggggggg gggggggsgr ggsggrgrgg
      781 kgcggcggca gccaatcaga gcggcgcgct ccgaaagttt ccttttatgg cgaggcggcg
      841 gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg ggagtcgctg cgcgctgcct
      901 tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg ccccggctct gactgaccgc
      961 gttactccca caggtgagcg ggcgggacgg cccttctcct ccgggctgta attagcgctt
     1021 ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa agccttgagg ggctccggga
     1081 gggccctttg tgcgggggga gcggctcggg gctgtccgcg gggggacggc tgccttcggg
     1141 ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac cggcggctct agagcctctg
     1201 ctaaccatgt tcatgccttc ttctttttcc tacagctcct gggcaacgtg ctggttattg
     1261 tgctgtctca tcattttggc aaagaattgg atcctctaga gtcgactccg gaataacttc
     1321 gtataggata ctttatacga agttatgcag aatggtagct ggattgtagc tgctattagc
     1381 aatatgaaac ctcttaataa cttcgtatag catacattat acgaagttat ggcgcgccgt
     1441 cgacttactt gtacagctcg tccatgccgt acaggaacag gtggtggcgg ccctcggagc
     1501 gctcgtactg ttccacgatg gtgtagtcct cgttgtggga ggtgatgtcc agcttggtgt
     1561 ccacgtagta gtagccgggc agttgcacgg gcttcttggc catgtagatg gtcttgaact
     1621 ccaccaggta gtggccgccg tccttcagct tcagggcctg gtggatctcg cccttcagca
     1681 cgccgtcgcg ggggtacagg cgctcggtgg aggcctccca gcccatggtc ttcttctgca
     1741 ttacggggcc gtcggggggg aagttggtgc cgcgcatctt caccttgtag atcagcgtgc
     1801 cgtcctgcag ggaggagtcc tgggtcacgg tcaccagacc gccgtcctcg aagttcatca
     1861 cgcgctccca cttgaagccc tcggggaagg acagcttctt gtaatcgggg atgtcggcgg
     1921 ggtgcttcac gtacgccttg gagccgtaca tgaactgggg ggacaggatg tcccaggcga
     1981 agggcagggg gccgcccttg gtcaccttca gcttggcggt ctgggtgccc tcgtaggggc
     2041 ggccctcgcc ctcgccctcg atctcgaact cgtggccgtt catggagccc tccatgcgca
     2101 ccttgaagcg catgaactct ttgatgacct cctcgccctt gctcaccatt ggcccgggat
     2161 tctcttcgac atcccctgct tgtttcaaca gggagaagtt agtggctccg cttccgttgt
     2221 cttcgaaccc accccccttg gatgcgcttt caagactgaa gatgtcagac ctgctttttg
     2281 ctttggactt cccccatgca gcgataccac ccacagctac caggcagcac aggagcactg
     2341 cagtgatcag catccacatg cggccgtgat tcctggctgg cagagcacgt ccaggagcag
     2401 gggcagtggg agcctctgtg ccgttgtccg tgggcaccgc ggggatcgcg ctggtcccgc
     2461 agccccactc gtcccgcccg tcgtcgcagt cggggtgtcc gtcgcacagc cagtcctgcg
     2521 ggtaacactc cccgtgggca ccgggcggct ccgagcagcg gaactgaccg ggggggcaac
     2581 gggacaaaga accgttaccg gacccgttac cggtcacgtt accgggcagc agcagcagca
     2641 gcagcgcggg cagcagccgc gccatggtgg cgctagcata acttcgtata aagtatccta
     2701 tacgaagtta tttgccttaa cccagaaatt atcactgtta ttctttagaa tggtgcaaag
     2761 aataacttcg tataatgtat gctatacgaa gttatgaatt cgatatcaag cttatcgata
     2821 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     2881 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     2941 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     3001 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     3061 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     3121 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     3181 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     3241 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     3301 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     3361 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgtc
     3421 gactcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     3481 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     3541 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     3601 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     3661 tatggcttct gaggcggaaa gaaccagctg gggctcgact agagcatggc tacgtagata
     3721 agtagcatgg cgggttaatc attaactaca aggaacccct agtgatggag ttggccactc
     3781 cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg
     3841 gcggcctcag tgagcgagcg agcgcgcaga gctttttgca aaagcctagg cctccaaaaa
     3901 agcctcctca ctacttctgg aatagctcag aggccgaggc ggcctcggcc tctgcataaa
     3961 taaaaaaaat tagtcagcca tggggcggag aatgggcgga actgggcgga gttaggggcg
     4021 ggatgggcgg agttaggggc gggactatgg ttgctgacta attgagatgc atgctttgca
     4081 tacttctgcc tgctggggag cctggggact ttccacacct ggttgctgac taattgagat
     4141 gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac cctaactgac
     4201 acacattcca cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     4261 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     4321 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     4381 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     4441 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     4501 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     4561 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     4621 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     4681 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     4741 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     4801 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     4861 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     4921 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     4981 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     5041 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     5101 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     5161 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     5221 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     5281 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     5341 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     5401 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     5461 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     5521 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     5581 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     5641 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     5701 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     5761 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     5821 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     5881 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     5941 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     6001 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     6061 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     6121 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     6181 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     6241 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     6301 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     6361 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     6421 ggcatcagag cagattgtac tgagagtgca ccattcgacg ctctccctta tgcgactcct
     6481 gcattaggaa gcagcccagt agtaggttga ggccgttgag caccgccgcc gcaaggaatg
     6541 gtgcatgcaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc accataccca
     6601 cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt
     6661 cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc
     6721 cggcgtagag gatctggcta gcgatgaccc tgctgattgg ttcgctgacc atttccgggt
     6781 gcgggacggc gttaccagaa actcagaagg ttcgtccaac caaaccgact ctgacggcag
     6841 tttacgagag agatgatagg gtctgcttca gtaagccaga tgctacacaa ttaggcttgt
     6901 acatattgtc gttagaacgc ggctacaatt aatacataac cttatgtatc atacacatac
     6961 gatttaggtg acactataga atacacggaa ttaa
//
