LOCUS       Exported                7321 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  An AAV vector that expresses engineered halorhodopsin 3.0 (fused to 
            iRFP) under the CaM Kinase IIa promoter..
ACCESSION   .
VERSION     .
KEYWORDS    pOTTC352 - pAAV CaMKIIa eNpHR3.0-iRFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7321)
  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 7321)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7321
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     promoter        160..1448
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     CDS             2440..3387
                     /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    3448..4036
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3501..3521)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3919..3930)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4068..4544
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4267..4286)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4584..4724
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4584..4713
                     /label=AAV2 ITR
     rep_origin      4799..5254
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4886..4905)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5096..5117
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5271..5290)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5390..5412
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5450..5468)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5536..5640
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5641..6501
                     /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(5859..5878)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6672..7260
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7161..7180
                     /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 cgcgtttaac attatggcct taggtcactt
      181 catctccatg gggttcttct tctgattttc tagaaaatga gatgggggtg cagagagctt
      241 cctcagtgac ctgcccaggg tcacatcaga aatgtcagag ctagaacttg aactcagatt
      301 actaatctta aattccatgc cttgggggca tgcaagtacg atatacagaa ggagtgaact
      361 cattagggca gatgaccaat gagtttagga aagaagagtc cagggcaggg tacatctaca
      421 ccacccgccc agccctgggt gagtccagcc acgttcacct cattatagtt gcctctctcc
      481 agtcctacct tgacgggaag cacaagcaga aactgggaca ggagccccag gagaccaaat
      541 cttcatggtc cctctgggag gatgggtggg gagagctgtg gcagaggcct caggaggggc
      601 cctgctgctc agtggtgaca gataggggtg agaaagcaga cagagtcatt ccgtcagcat
      661 tctgggtctg tttggtactt cttctcacgc taaggtggcg gtgtgatatg cacaatggct
      721 aaaaagcagg gagagctgga aagaaacaag gacagagaca gaggccaagt caaccagacc
      781 aattcccaga ggaagcaaag aaaccattac agagactaca agggggaagg gaaggagaga
      841 tgaattagct tcccctgtaa accttagaac ccagctgttg ccagggcaac ggggcaatac
      901 ctgtctcttc agaggagatg aagttgccag ggtaactaca tcctgtcttt ctcaaggacc
      961 atcccagaat gtggcaccca ctagccgtta ccatagcaac tgcctctttg ccccacttaa
     1021 tcccatcccg tctgttaaaa gggccctata gttggaggtg ggggaggtag gaagagcgat
     1081 gatcacttgt ggactaagtt tgttcgcatc cccttctcca accccctcag tacatcaccc
     1141 tgggggaaca gggtccactt gctcctgggc ccacacagtc ctgcagtatt gtgtatataa
     1201 ggccagggca aagaggagca ggttttaaag tgaaaggcag gcaggtgttg gggaggcagt
     1261 taccggggca acgggaacag ggcgtttcgg aggtggttgc catggggacc tggatgctga
     1321 cgaaggctcg cgaggctgtg agcagccaca gtgccctgct cagaagcccc aagctcgtca
     1381 gtcaagccgg ttctccgttt gcactcagga gcacgggcag gcgagtggcc cctagttctg
     1441 ggggcagctc tagagcggta ccggatcccc gggtaccggt aggccaccat gacagagacc
     1501 ctgcctcccg tgaccgagag tgccgtggcc cttcaagccg aggttaccca aagggagttg
     1561 ttcgagttcg tgctgaacga ccctttgctt gcaagcagtc tctatatcaa catcgcactt
     1621 gcaggactga gtatactgct gttcgttttt atgacccgag gactcgatga tccacgggca
     1681 aaacttattg ctgtgtcaac catccttgtg cctgtcgtca gcattgcctc ctacactgga
     1741 ttggcgagcg gcctgacaat ttccgttctt gaaatgccag cgggccattt tgcagaaggc
     1801 agctcagtga tgctgggagg agaagaggta gatggtgtag tcaccatgtg gggacggtat
     1861 ctcacctggg cactttccac gcccatgatt ctcctcgctc tgggtctcct ggccggaagc
     1921 aatgctacaa agctcttcac agctatcact ttcgatatcg ctatgtgcgt gactggcctt
     1981 gccgcggccc tgactacctc ctcccacctc atgagatggt tctggtacgc tatcagttgt
     2041 gcatgctttc tggtggtctt gtatatcctg ctggtggagt gggcacagga cgccaaagcc
     2101 gcgggaaccg ctgacatgtt caataccctg aagctgttga cagtagtgat gtggctgggg
     2161 tatccaattg tgtgggctct tggagtcgag ggtatcgcgg tgttgcccgt tggggtgacg
     2221 agctggggat attctttcct ggatatcgtg gcaaagtaca ttttcgcatt cttgctcctg
     2281 aactatctga cgtcaaacga atctgtcgtg tccggcagca ttttggatgt tccatctgct
     2341 tctgggaccc cggctgatga tgcggccgcc aagagcagga tcaccagcga gggcgagtac
     2401 atccccctgg accagatcga catcaacgtg gtgactagta tggctgaagg atccgtcgcc
     2461 aggcagcctg acctcttgac ctgcgacgat gagccgatcc atatccccgg tgccatccaa
     2521 ccgcatggac tgctgctcgc cctcgccgcc gacatgacga tcgttgccgg cagcgacaac
     2581 cttcccgaac tcaccggact ggcgatcggc gccctgatcg gccgctctgc ggccgatgtc
     2641 ttcgactcgg agacgcacaa ccgtctgacg atcgccttgg ccgagcccgg ggcggccgtc
     2701 ggagcaccga tcactgtcgg cttcacgatg cgaaaggacg caggcttcat cggctcctgg
     2761 catcgccatg atcagctcat cttcctcgag ctcgagcctc cccagcggga cgtcgccgag
     2821 ccgcaggcgt tcttccgccg caccaacagc gccatccgcc gcctgcaggc cgccgaaacc
     2881 ttggaaagcg cctgcgccgc cgcggcgcaa gaggtgcgga agattaccgg cttcgatcgg
     2941 gtgatgatct atcgcttcgc ctccgacttc agcggcgaag tgatcgcaga ggatcggtgc
     3001 gccgaggtcg agtcaaaact aggcctgcac tatcctgcct caaccgtgcc ggcgcaggcc
     3061 cgtcggctct ataccatcaa cccggtacgg atcattcccg atatcaatta tcggccggtg
     3121 ccggtcaccc cagacctcaa tccggtcacc gggcggccga ttgatcttag cttcgccatc
     3181 ctgcgcagcg tctcgcccgt ccatctggaa ttcatgcgca acataggcat gcacggcacg
     3241 atgtcgatct cgattttgcg cggcgagcga ctgtggggat tgatcgtttg ccatcaccga
     3301 acgccgtact acgtcgatct cgatggccgc caagcctgcg agctagtcgc ccaggttctg
     3361 gcctggcaga tcggcgtgat ggaagagctg tacaagttct gctacgagaa cgaggtgtaa
     3421 tgagaattcg atatcaagct tatcgataat caacctctgg attacaaaat ttgtgaaaga
     3481 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     3541 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     3601 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     3661 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     3721 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     3781 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     3841 aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
     3901 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     3961 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
     4021 tgggccgcct ccccgcatcg ataccgagcg ctgctcgaga gatctacggg tggcatccct
     4081 gtgacccctc cccagtgcct ctcctggccc tggaagttgc cactccagtg cccaccagcc
     4141 ttgtcctaat aaaattaagt tgcatcattt tgtctgacta ggtgtccttc tataatatta
     4201 tggggtggag gggggtggta tggagcaagg ggcaagttgg gaagacaacc tgtagggcct
     4261 gcggggtcta ttgggaacca agctggagtg cagtggcaca atcttggctc actgcaatct
     4321 ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc cgagttgttg ggattccagg
     4381 catgcatgac caggctcagc taatttttgt ttttttggta gagacggggt ttcaccatat
     4441 tggccaggct ggtctccaac tcctaatctc aggtgatcta cccaccttgg cctcccaaat
     4501 tgctgggatt acaggcgtga accactgctc ccttccctgt ccttctgatt ttgtaggtaa
     4561 ccacgtgcgg accgagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct
     4621 gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc
     4681 ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta
     4741 ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac
     4801 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     4861 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     4921 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     4981 gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca
     5041 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     5101 ctcttgttcc aaactggaac aacactcaac cctatctcgg gctattcttt tgatttataa
     5161 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     5221 gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc
     5281 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga
     5341 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     5401 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata
     5461 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     5521 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     5581 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     5641 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     5701 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     5761 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     5821 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     5881 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     5941 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     6001 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     6061 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     6121 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     6181 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     6241 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     6301 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     6361 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     6421 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     6481 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     6541 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     6601 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     6661 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     6721 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     6781 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
     6841 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     6901 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     6961 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     7021 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     7081 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     7141 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     7201 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     7261 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     7321 t
//
