LOCUS       sequence-68472-e        6690 bp    DNA     circular SYN 14-APR-2025
DEFINITION  An AAV vector that expresses iRFP under the c-fos promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6690)
  AUTHORS   Richie CT, Whitaker LR, Whitaker KW, Necarsulmer J, Baldwin HA, 
            Zhang Y, Fortuno L, Hinkle JJ, Koivula P, Henderson MJ, Sun W, Wang 
            K, Smith JC, Pickel J, Ji N, Hope BT, Harvey BK
  TITLE     Near-infrared fluorescent protein iRFP713 as a reporter protein for 
            optogenetic vectors, a transgenic Cre-reporter rat, and other 
            neuronal studies.
  JOURNAL   J Neurosci Methods. 2017 Jun 1;284:1-14. doi: 
            10.1016/j.jneumeth.2017.03.020. Epub 2017 Apr 2.
  PUBMED    28380331
REFERENCE   2  (bases 1 to 6690)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Neurosci 
            Methods."; date: "2017-06-1"; volume: "284:1-14. doi"; pages: " 
            10.1016/j.jneumeth.2017.03.020. Epub 2017 Apr 2"
COMMENT     Sequence Label: pOTTC475 - pAAV c-fos iRFP
FEATURES             Location/Qualifiers
     source          1..6690
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     CDS             complement(856..867)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             1833..2780
                     /codon_start=1
                     /product="phytochrome-based near-infrared fluorescent 
                     protein, originally termed iRFP (Filonov et al., 2011)"
                     /label=iRFP713
                     /note="name was changed to iRFP713 (Shcherbakova and 
                     Verkhusha, 2013)"
                     /translation="MAEGSVARQPDLLTCDDEPIHIPGAIQPHGLLLALAADMTIVAGS
                     DNLPELTGLAIGALIGRSAADVFDSETHNRLTIALAEPGAAVGAPITVGFTMRKDAGFI
                     GSWHRHDQLIFLELEPPQRDVAEPQAFFRRTNSAIRRLQAAETLESACAAAAQEVRKIT
                     GFDRVMIYRFASDFSGEVIAEDRCAEVESKLGLHYPASTVPAQARRLYTINPVRIIPDI
                     NYRPVPVTPDLNPVTGRPIDLSFAILRSVSPVHLEFMRNIGMHGTMSISILRGERLWGL
                     IVCHHRTPYYVDLDGRQACELVAQVLAWQIGVMEE"
     misc_feature    2817..3405
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2870..2890)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3288..3299)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3437..3913
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3636..3655)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3953..4093
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3953..4082
                     /label=AAV2 ITR
     rep_origin      4168..4623
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4255..4274)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4465..4486
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4640..4659)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4759..4781
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4819..4837)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4905..5009
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5010..5078,5079..5870)
                     /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(5228..5247)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6041..6629
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6530..6549
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtaagct ttcctttagg aacagaggct
      181 tcgagccttt aaggctgcgt acttgcttct cctaatacca gagactcaaa aaaaaaaaaa
      241 aagttccaga ttgctggaca atgacccggg tctcatccct tgaccctggg aaccgggtcc
      301 acattgaatc aggtgcgaat gttcgctcgc cttctctgcc tttcccgcct cccctccccc
      361 ggccgcggcc ccggttcccc ccctgcgctg caccctcaga gttggctgca gccggcgagc
      421 tgttcccgtc aatccctccc tcctttacac aggatgtcca tattaggaca tctgcgtcag
      481 caggtttcca cggccggtcc ctgttgttct ggggggggga ccatctccga aatcctacac
      541 gcggaaggtc taggagaccc cctaagatcc caaatgtgaa cactcatagg tgaaagatgt
      601 atgccaagac gggggttgaa agcctggggc gtagagttga cgacagagcg cccgcagagg
      661 gccttggggc gcgcttcccc ccccttccag ttccgcccag tgacgtagga agtccatcca
      721 ttcacagcgc ttctataaag gcgccagctg aggcgcctac tactccaacc gcgactgcag
      781 cgagcaactg agaagactgg atagagccgg cggttccgcg aacgagcagt gaccgcgctc
      841 ccacccagct ctgctctgca gctcccacca gtgtctaccc ctggacccct tgccgggctt
      901 tccccaaact tcgaccatga tgttctcggg tttcaacgcc gactacgagg cgtcatcctc
      961 ccgctgcagt agcgcctccc cggccgggga cagcctttcc tactaccatt ccccagccga
     1021 ctccttctcc agcatgggct ctcctgtcaa cacacaggtg agtttggctt tgtgtagccg
     1081 ccaggtccgc gctgagggtc gccgtggagg agacactggg gtgtgactcg caggggcggg
     1141 ggggtcttcc tttttcgctc tggagggaga ctggcgcggt cagagcagcc ttagcctggg
     1201 aacccaggac ttgtctgagc gcgtgcacac ttgtcatagt aagacttagt gaccccttcc
     1261 cgcgcggcag gtttattctg agtggcctgc ctgcattctt ctctcggccg acttgtttct
     1321 gagatcagcc ggggccaaca agtctcgagc aaagagtcgc taactagagt ttgggaggcg
     1381 gcaaaccgcg gcaatccccc ctcccggggc agcctggagc agggaggagg gnggagggag
     1441 gagggtgctg cgggcgggtg tgtaaggcag tttcattgat aaaaagcgag ttcattctgg
     1501 agactccgga gcagcgcctg cgtcagcgca gacgtcaggg atatttataa caaaccccct
     1561 ttcgagcgag tgatgccgaa gggataacgg gaacgcagca gtaggatgga ggggaaaggc
     1621 tgcgctgcgg aattcaaggg aggatattgg gagagctttt atctccgatg aggtgcatac
     1681 aggaagacat aagcagtctc tgaccggaat gcttctctct ccctgcttca tgcgacacta
     1741 gggccacttg ctccacctgt gtctggaacc tcctcgctca cctccgcttt cctctttttg
     1801 ttttgtttca ggacttttgc gcagatctct ccatggctga aggatccgtc gccaggcagc
     1861 ctgacctctt gacctgcgac gatgagccga tccatatccc cggtgccatc caaccgcatg
     1921 gactgctgct cgccctcgcc gccgacatga cgatcgttgc cggcagcgac aaccttcccg
     1981 aactcaccgg actggcgatc ggcgccctga tcggccgctc tgcggccgat gtcttcgact
     2041 cggagacgca caaccgtctg acgatcgcct tggccgagcc cggggcggcc gtcggagcac
     2101 cgatcactgt cggcttcacg atgcgaaagg acgcaggctt catcggctcc tggcatcgcc
     2161 atgatcagct catcttcctc gagctcgagc ctccccagcg ggacgtcgcc gagccgcagg
     2221 cgttcttccg ccgcaccaac agcgccatcc gccgcctgca ggccgccgaa accttggaaa
     2281 gcgcctgcgc cgccgcggcg caagaggtgc ggaagattac cggcttcgat cgggtgatga
     2341 tctatcgctt cgcctccgac ttcagcggcg aagtgatcgc agaggatcgg tgcgccgagg
     2401 tcgagtcaaa actaggcctg cactatcctg cctcaaccgt gccggcgcag gcccgtcggc
     2461 tctataccat caacccggta cggatcattc ccgatatcaa ttatcggccg gtgccggtca
     2521 ccccagacct caatccggtc accgggcggc cgattgatct tagcttcgcc atcctgcgca
     2581 gcgtctcgcc cgtccatctg gaattcatgc gcaacatagg catgcacggc acgatgtcga
     2641 tctcgatttt gcgcggcgag cgactgtggg gattgatcgt ttgccatcac cgaacgccgt
     2701 actacgtcga tctcgatggc cgccaagcct gcgagctagt cgcccaggtt ctggcctggc
     2761 agatcggcgt gatggaagag ttgtacaagt aagaattcga tatcaagctt atcgataatc
     2821 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     2881 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     2941 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     3001 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     3061 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     3121 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     3181 gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct
     3241 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     3301 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     3361 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcatcga taccgagcgc
     3421 tgctcgagag atctacgggt ggcatccctg tgacccctcc ccagtgcctc tcctggccct
     3481 ggaagttgcc actccagtgc ccaccagcct tgtcctaata aaattaagtt gcatcatttt
     3541 gtctgactag gtgtccttct ataatattat ggggtggagg ggggtggtat ggagcaaggg
     3601 gcaagttggg aagacaacct gtagggcctg cggggtctat tgggaaccaa gctggagtgc
     3661 agtggcacaa tcttggctca ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc
     3721 tcagcctccc gagttgttgg gattccaggc atgcatgacc aggctcagct aatttttgtt
     3781 tttttggtag agacggggtt tcaccatatt ggccaggctg gtctccaact cctaatctca
     3841 ggtgatctac ccaccttggc ctcccaaatt gctgggatta caggcgtgaa ccactgctcc
     3901 cttccctgtc cttctgattt tgtaggtaac cacgtgcgga ccgagcggcc gcaggaaccc
     3961 ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga
     4021 ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc
     4081 agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     4141 caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg
     4201 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     4261 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     4321 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     4381 atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     4441 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     4501 ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     4561 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa
     4621 ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac
     4681 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     4741 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     4801 aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     4861 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     4921 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     4981 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     5041 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     5101 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     5161 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     5221 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     5281 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     5341 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     5401 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     5461 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     5521 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     5581 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     5641 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     5701 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     5761 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     5821 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     5881 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     5941 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     6001 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     6061 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     6121 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     6181 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     6241 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     6301 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     6361 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     6421 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     6481 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     6541 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     6601 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     6661 tggccttttg ctggcctttt gctcacatgt
//
