LOCUS       Exported                7957 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Cre- dependent expression of Sun1GFP for nuclei isolation.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-Ef1a-DIO-Sun1GFP-WPRE-pA
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7957)
  AUTHORS   Lawler AJ, Brown AR, Bouchard RS, Toong N, Kim Y, Velraj N, Fox G, 
            Kleyman M, Kang B, Gittis AH, Pfenning AR
  TITLE     Cell type-specific oxidative stress genomic signatures in the globus
            pallidus of dopamine depleted mice.
  JOURNAL   J Neurosci. 2020 Nov 10. pii: JNEUROSCI.1634-20.2020. doi: 
            10.1523/JNEUROSCI.1634-20.2020.
  PUBMED    33188066
REFERENCE   2  (bases 1 to 7957)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7957
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        231..1409
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          462..1400
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1357..1377
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    1440..1473
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites (Lee and 
                     Saito, 1988)."
     protein_bind    1524..1557
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     CDS             complement(1569..2282)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(1609..1629)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     2224..2252
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     protein_bind    complement(3942..3975)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites (Lee and 
                     Saito, 1988)."
     protein_bind    complement(4026..4059)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    4084..4672
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4137..4157)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4555..4566)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4704..5180
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4903..4922)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   5220..5360
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5220..5349
                     /label=AAV2 ITR
     rep_origin      5435..5890
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5522..5541)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5732..5753
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5907..5926)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6026..6048
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6086..6104)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6172..6276
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6277..7137
                     /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(6495..6514)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7308..7896
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7797..7816
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtaagctt tgcaaagatg gataaagttt taaacagaga
      181 ggaatctttg cagctaatgg accttctagg tcttgaaagg agtgggaatt ggctccggtg
      241 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
      301 gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
      361 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
      421 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt
      481 cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct
      541 ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga
      601 ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct
      661 ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt
      721 ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc
      781 ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg
      841 acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac
      901 cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc
      961 cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag
     1021 cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct
     1081 cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg
     1141 tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga
     1201 gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc
     1261 acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg
     1321 aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa
     1381 gtttttttct tccatttcag gtgtcgtgag gtaccggatc ctctagagtc gactccggaa
     1441 taacttcgta taggatactt tatacgaagt tatgcagaat ggtagctgga ttgtagctgc
     1501 tattagcaat atgaaacctc ttaataactt cgtatagcat acattatacg aagttatggc
     1561 gcgccttatt tgtagagctc atccatgcca tgtgtaatcc cagcagcagt tacaaactca
     1621 agaaggacca tgtggtcacg cttttcgttg ggatctttcg aaaggacaga ttgtgtcgac
     1681 aggtaatggt tgtctggtaa aaggacaggg ccatcgccaa ttggagtatt ttgttgataa
     1741 tggtctgcta gttgaacgga accatcttca acgttgtggc gaattttgaa gttagctttg
     1801 attccattct tttgtttgtc tgccgtgatg tatacattgt gtgagttaaa gttgtactcg
     1861 agtttgtgtc cgagaatgtt tccatcttct ttaaaatcaa taccttttaa ctcgatacga
     1921 ttaacaaggg tatcaccttc aaacttgact tcagcacgcg tcttgtaggt cccgtcatct
     1981 ttgaaagata tagtgcgttc ctgtacataa ccttcgggca tggcactctt gaaaaagtca
     2041 tgccgtttca tgtgatccgg ataacgggaa aagcattgaa caccataggt cagagtagtg
     2101 acaagtgttg gccatggaac aggtagtttt ccagtagtgc aaataaattt aagggtgagt
     2161 tttccgtttg tagcatcacc ttcaccctct ccacggacag aaaatttgtg cccattaaca
     2221 tcaccatcta attcaacaag aattgggaca actccagtga aaagttcttc tcctttgctc
     2281 atctggatgg gctctccgtg gactcggaac cggtagaggc aagtgtactc agggtggccc
     2341 cagttggaca ggacccggag ctctactatc tggaaggctt ggtcaggtct ttccagtgtg
     2401 tggaacatct ggagtgagtc tccttcctgg tcataggtga accgtcccag aggctgcccc
     2461 tcttcttgat actccgtttc cagtccatag actgcaaagt ctttgggggc actggagatg
     2521 ttaccagtgg gtgatagtgt ctttggaatg tgttccatgg tgaatgtggt tgggtagatc
     2581 ttcatggaca accgcaccac caggtacccc tgggaacctt tgaacgccca gcaattccct
     2641 gggtagatgt cgggctggat caccactcga ggtgactgtg agaagtacca cagtgggacc
     2701 ccaaacaggc tcagcagtgc cgtcttggtc tcataggtct cagagcaccg agtgcttagg
     2761 atgctgccac ctccagactc cagagcaaag tccaccatcc ccgtcttgtc ttgggagtac
     2821 agcttcagag cattgttcac aatgatatgt gcttgcgctt ctgtgattcc tgaaatccct
     2881 gcctgattca cggcagacac aatagcctcg gatgtcgggg cctgtcctgt cacggtgatg
     2941 tggtgtgtga tattctgcag cagtttcagc tcaaggtcat gtaagagcac ctgcagctca
     3001 tccttgctca cgaaccgaga agaaagcttc tctaatagcc actcgaggga accgccctgc
     3061 tgatcctcag agaacatgag ctgcacagtc tcctggatgc gggccccagc ctgctcacag
     3121 ctggtcttca gatgctgcca gtctgacagc tgtgacttca acaggttcag ttccagttct
     3181 aggtgctgca cacggtcaag gaggggctgc tcttcatccc tgctgcctgc tttcagcttg
     3241 gtttcttcca gctccttctg gatagcctca gatttttctg tcagttttct aagaacatct
     3301 tccaagttgg agagacgcac ttcatggtca tggtggaaag tcatgaagtc agtcttagca
     3361 tcaggtggtt ctatggtgcc catctcctgc aggtgctgtc caaccacatt cttgacccac
     3421 agtgacagcc cttccctgcc gtcatccact tggtctaccc gtatctgtag tttctgaagc
     3481 aggactgtca gctctgcaag tctctcatca tggttgtggc actgcgcact caaagaagct
     3541 accttctgcc ccatgtcact ctcctgaggc cactgggaag ccttgtgatc aacctttgga
     3601 ggtgggctcg ggggaagagg gccaggtcta tgcatgccct tggaatcgtc caccctctgt
     3661 gttggctgca tggccgtcca gttcagcact ggtaggagtg agaagaagtt tccctggccc
     3721 cacagggaga caccagcacc tagtaaaagt aggagtggga ggagcaagac aaaaaccttg
     3781 caaatatttc gaaggcacct ggtaagaaga aagacattca gccaagaaat caaagtaaca
     3841 aattggtacc agccgctccc tagccaccag aaggttcccg aggctgcctt ccgtggtttg
     3901 agagtcctgt ctctggagta caccctggag gtcatgctag cataacttcg tataaagtat
     3961 cctatacgaa gttatttgcc ttaacccaga aattatcact gttattcttt agaatggtgc
     4021 aaagaataac ttcgtataat gtatgctata cgaagttatg aattcgatat caagcttatc
     4081 gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     4141 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     4201 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     4261 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     4321 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     4381 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     4441 ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg
     4501 ctcgcctatg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     4561 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     4621 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcatcgatac
     4681 cgagcgctgc tcgagagatc tacgggtggc atccctgtga cccctcccca gtgcctctcc
     4741 tggccctgga agttgccact ccagtgccca ccagccttgt cctaataaaa ttaagttgca
     4801 tcattttgtc tgactaggtg tccttctata atattatggg gtggaggggg gtggtatgga
     4861 gcaaggggca agttgggaag acaacctgta gggcctgcgg ggtctattgg gaaccaagct
     4921 ggagtgcagt ggcacaatct tggctcactg caatctccgc ctcctgggtt caagcgattc
     4981 tcctgcctca gcctcccgag ttgttgggat tccaggcatg catgaccagg ctcagctaat
     5041 ttttgttttt ttggtagaga cggggtttca ccatattggc caggctggtc tccaactcct
     5101 aatctcaggt gatctaccca ccttggcctc ccaaattgct gggattacag gcgtgaacca
     5161 ctgctccctt ccctgtcctt ctgattttgt aggtaaccac gtgcggaccg agcggccgca
     5221 ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc
     5281 cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg
     5341 agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     5401 tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc
     5461 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cttagcgccc
     5521 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     5581 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     5641 aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     5701 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     5761 ctcaactcta tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggtctat
     5821 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     5881 tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     5941 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     6001 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     6061 tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     6121 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     6181 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     6241 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     6301 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     6361 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     6421 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     6481 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     6541 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     6601 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     6661 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     6721 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     6781 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     6841 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     6901 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     6961 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     7021 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     7081 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     7141 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     7201 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     7261 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     7321 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     7381 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     7441 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     7501 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     7561 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     7621 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     7681 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     7741 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     7801 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     7861 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     7921 cggttcctgg ccttttgctg gccttttgct cacatgt
//
