LOCUS       Exported                6534 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  An AAV vector that expresses nuclear localized eYFP under control of
            the c-fos promoter..
ACCESSION   .
VERSION     .
KEYWORDS    pOTTC589	- pAAV c-fos Nuc-eYFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6534)
  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 6534)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6534
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             complement(497..508)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             1474..2190
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1519..1540)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1780..1799)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2127..2148
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2215..2235
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2239..2259
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2263..2283
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    2314..2902
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2367..2387)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2785..2796)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2934..3410
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3133..3152)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3450..3590
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3450..3579
                     /label=AAV2 ITR
     rep_origin      3665..4120
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3752..3771)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3962..3983
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4137..4156)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4256..4278
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4316..4334)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4402..4506
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4507..5367
                     /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(4725..4744)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5538..6126
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6027..6046
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   6188..6317
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 cggccgcggc cccggttccc cccctgcgct gcaccctcag agttggctgc agccggcgag
       61 ctgttcccgt caatccctcc ctcctttaca caggatgtcc atattaggac atctgcgtca
      121 gcaggtttcc acggccggtc cctgttgttc tggggggggg accatctccg aaatcctaca
      181 cgcggaaggt ctaggagacc ccctaagatc ccaaatgtga acactcatag gtgaaagatg
      241 tatgccaaga cgggggttga aagcctgggg cgtagagttg acgacagagc gcccgcagag
      301 ggccttgggg cgcgcttccc cccccttcca gttccgccca gtgacgtagg aagtccatcc
      361 attcacagcg cttctataaa ggcgccagct gaggcgccta ctactccaac cgcgactgca
      421 gcgagcaact gagaagactg gatagagccg gcggttccgc gaacgagcag tgaccgcgct
      481 cccacccagc tctgctctgc agctcccacc agtgtctacc cctggacccc ttgccgggct
      541 ttccccaaac ttcgaccatg atgttctcgg gtttcaacgc cgactacgag gcgtcatcct
      601 cccgctgcag tagcgcctcc ccggccgggg acagcctttc ctactaccat tccccagccg
      661 actccttctc cagcatgggc tctcctgtca acacacaggt gagtttggct ttgtgtagcc
      721 gccaggtccg cgctgagggt cgccgtggag gagacactgg ggtgtgactc gcaggggcgg
      781 gggggtcttc ctttttcgct ctggagggag actggcgcgg tcagagcagc cttagcctgg
      841 gaacccagga cttgtctgag cgcgtgcaca cttgtcatag taagacttag tgaccccttc
      901 ccgcgcggca ggtttattct gagtggcctg cctgcattct tctctcggcc gacttgtttc
      961 tgagatcagc cggggccaac aagtctcgag caaagagtcg ctaactagag tttgggaggc
     1021 ggcaaaccgc ggcaatcccc cctcccgggg cagcctggag cagggaggag ggaggaggga
     1081 ggagggtgct gcgggcgggt gtgtaaggca gtttcattga taaaaagcga gttcattctg
     1141 gagactccgg agcagcgcct gcgtcagcgc agacgtcagg gatatttata acaaaccccc
     1201 tttcgagcga gtgatgccga agggataacg ggaacgcagc agtaggatgg aggggaaagg
     1261 ctgcgctgcg gaattcaagg gaggatattg ggagagcttt tatctccgat gaggtgcata
     1321 caggaagaca taagcagtct ctgaccggaa tgcttctctc tccctgcttc atgcgacact
     1381 agggccactt gctccacctg tgtctggaac ctcctcgctc acctccgctt tcctcttttt
     1441 gttttgtttc aggacttttg cgcagatctc tccatggtga gcaagggcga ggagctgttc
     1501 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     1561 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     1621 accaccggca agctgcccgt gccctggccc accctcgtga ccaccttcgg ctacggcctg
     1681 cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     1741 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     1801 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     1861 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     1921 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     1981 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     2041 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gctaccagtc cgccctgagc
     2101 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     2161 atcactctcg gcatggacga gctgtacaag tccggcctgc gcagccgcgc cgacccaaaa
     2221 aagaagcgca aggtagaccc caagaagaag cgcaaggtag atccaaaaaa gaagagaaag
     2281 gtatgatagg aattcgatat caagcttatc gataatcaac ctctggatta caaaatttgt
     2341 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     2401 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     2461 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     2521 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     2581 ctcctttccg gaactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     2641 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     2701 tcggggaaat catcgtcctt tccttggctg ctcgcctatg ttgccacctg gattctgcgc
     2761 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     2821 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     2881 tccctttggg ccgcctcccc gcatcgatac cgagcgctgc tcgagagatc tacgggtggc
     2941 atccctgtga cccctcccca gtgcctctcc tggccctgga agttgccact ccagtgccca
     3001 ccagccttgt cctaataaaa ttaagttgca tcattttgtc tgactaggtg tccttctata
     3061 atattatggg gtggaggggg gtggtatgga gcaaggggca agttgggaag acaacctgta
     3121 gggcctgcgg ggtctattgg gaaccaagct ggagtgcagt ggcacaatct tggctcactg
     3181 caatctccgc ctcctgggtt caagcgattc tcctgcctca gcctcccgag ttgttgggat
     3241 tccaggcatg catgaccagg ctcagctaat ttttgttttt ttggtagaga cggggtttca
     3301 ccatattggc caggctggtc tccaactcct aatctcaggt gatctaccca ccttggcctc
     3361 ccaaattgct gggattacag gcgtgaacca ctgctccctt ccctgtcctt ctgattttgt
     3421 aggtaaccac gtgcggaccg agcggccgca ggaaccccta gtgatggagt tggccactcc
     3481 ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg
     3541 ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat
     3601 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca
     3661 tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     3721 accgctacac ttgccagcgc cttagcgccc gctcctttcg ctttcttccc ttcctttctc
     3781 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     3841 tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt
     3901 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     3961 agtggactct tgttccaaac tggaacaaca ctcaactcta tctcgggcta ttcttttgat
     4021 ttataaggga ttttgccgat ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa
     4081 tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca
     4141 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
     4201 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     4261 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc
     4321 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     4381 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     4441 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     4501 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     4561 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     4621 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     4681 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     4741 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     4801 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     4861 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     4921 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     4981 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     5041 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     5101 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     5161 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     5221 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     5281 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     5341 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     5401 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     5461 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     5521 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     5581 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     5641 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     5701 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     5761 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     5821 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     5881 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     5941 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     6001 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     6061 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     6121 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     6181 cacatgtcct gcaggcagct gcgcgctcgc tcgctcactg aggccgcccg ggcgtcgggc
     6241 gacctttggt cgcccggcct cagtgagcga gcgagcgcgc agagagggag tggccaactc
     6301 catcactagg ggttcctgcg gccgcacgcg taagctttcc tttaggaaca gaggcttcga
     6361 gcctttaagg ctgcgtactt gcttctccta ataccagaga ctcaaaaaaa aaaaaaagtt
     6421 ccagattgct ggacaatgac ccgggtctca tcccttgacc ctgggaaccg ggtccacatt
     6481 gaatcaggtg cgaatgttcg ctcgccttct ctgcctttcc cgcctcccct cccc
//
