LOCUS       Exported                7967 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 7967)
  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 7967)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7967
                     /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        242..1419
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          473..1410
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1367..1387
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    1450..1483
                     /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    1534..1567
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     CDS             complement(1579..2292)
                     /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(1619..1639)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     2234..2262
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     protein_bind    complement(3952..3985)
                     /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(4036..4069)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    4094..4682
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4147..4167)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4565..4576)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4714..5190
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4913..4932)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   5230..5370
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5230..5359
                     /label=AAV2 ITR
     rep_origin      5445..5900
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5532..5551)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5742..5763
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5917..5936)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6036..6058
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6096..6114)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6182..6286
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6287..7147
                     /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(6505..6524)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7318..7906
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7807..7826
                     /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 ttgcaaagat ggataaagtt
      181 ttaaacagag aggaatcttt gcagctaatg gaccttctag gtcttgaaag gagtgggaat
      241 tggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
      301 gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag
      361 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc
      421 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc
      481 cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat
      541 tacttccact ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      601 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      661 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      721 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      781 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      841 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      901 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      961 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
     1021 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
     1081 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
     1141 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
     1201 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1261 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1321 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1381 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gtaccggatc ctctagagtc
     1441 gactccggaa taacttcgta taggatactt tatacgaagt tatgcagaat ggtagctgga
     1501 ttgtagctgc tattagcaat atgaaacctc ttaataactt cgtatagcat acattatacg
     1561 aagttatggc gcgccttatt tgtagagctc atccatgcca tgtgtaatcc cagcagcagt
     1621 tacaaactca agaaggacca tgtggtcacg cttttcgttg ggatctttcg aaaggacaga
     1681 ttgtgtcgac aggtaatggt tgtctggtaa aaggacaggg ccatcgccaa ttggagtatt
     1741 ttgttgataa tggtctgcta gttgaacgga accatcttca acgttgtggc gaattttgaa
     1801 gttagctttg attccattct tttgtttgtc tgccgtgatg tatacattgt gtgagttaaa
     1861 gttgtactcg agtttgtgtc cgagaatgtt tccatcttct ttaaaatcaa taccttttaa
     1921 ctcgatacga ttaacaaggg tatcaccttc aaacttgact tcagcacgcg tcttgtaggt
     1981 cccgtcatct ttgaaagata tagtgcgttc ctgtacataa ccttcgggca tggcactctt
     2041 gaaaaagtca tgccgtttca tgtgatccgg ataacgggaa aagcattgaa caccataggt
     2101 cagagtagtg acaagtgttg gccatggaac aggtagtttt ccagtagtgc aaataaattt
     2161 aagggtgagt tttccgtttg tagcatcacc ttcaccctct ccacggacag aaaatttgtg
     2221 cccattaaca tcaccatcta attcaacaag aattgggaca actccagtga aaagttcttc
     2281 tcctttgctc atctggatgg gctctccgtg gactcggaac cggtagaggc aagtgtactc
     2341 agggtggccc cagttggaca ggacccggag ctctactatc tggaaggctt ggtcaggtct
     2401 ttccagtgtg tggaacatct ggagtgagtc tccttcctgg tcataggtga accgtcccag
     2461 aggctgcccc tcttcttgat actccgtttc cagtccatag actgcaaagt ctttgggggc
     2521 actggagatg ttaccagtgg gtgatagtgt ctttggaatg tgttccatgg tgaatgtggt
     2581 tgggtagatc ttcatggaca accgcaccac caggtacccc tgggaacctt tgaacgccca
     2641 gcaattccct gggtagatgt cgggctggat caccactcga ggtgactgtg agaagtacca
     2701 cagtgggacc ccaaacaggc tcagcagtgc cgtcttggtc tcataggtct cagagcaccg
     2761 agtgcttagg atgctgccac ctccagactc cagagcaaag tccaccatcc ccgtcttgtc
     2821 ttgggagtac agcttcagag cattgttcac aatgatatgt gcttgcgctt ctgtgattcc
     2881 tgaaatccct gcctgattca cggcagacac aatagcctcg gatgtcgggg cctgtcctgt
     2941 cacggtgatg tggtgtgtga tattctgcag cagtttcagc tcaaggtcat gtaagagcac
     3001 ctgcagctca tccttgctca cgaaccgaga agaaagcttc tctaatagcc actcgaggga
     3061 accgccctgc tgatcctcag agaacatgag ctgcacagtc tcctggatgc gggccccagc
     3121 ctgctcacag ctggtcttca gatgctgcca gtctgacagc tgtgacttca acaggttcag
     3181 ttccagttct aggtgctgca cacggtcaag gaggggctgc tcttcatccc tgctgcctgc
     3241 tttcagcttg gtttcttcca gctccttctg gatagcctca gatttttctg tcagttttct
     3301 aagaacatct tccaagttgg agagacgcac ttcatggtca tggtggaaag tcatgaagtc
     3361 agtcttagca tcaggtggtt ctatggtgcc catctcctgc aggtgctgtc caaccacatt
     3421 cttgacccac agtgacagcc cttccctgcc gtcatccact tggtctaccc gtatctgtag
     3481 tttctgaagc aggactgtca gctctgcaag tctctcatca tggttgtggc actgcgcact
     3541 caaagaagct accttctgcc ccatgtcact ctcctgaggc cactgggaag ccttgtgatc
     3601 aacctttgga ggtgggctcg ggggaagagg gccaggtcta tgcatgccct tggaatcgtc
     3661 caccctctgt gttggctgca tggccgtcca gttcagcact ggtaggagtg agaagaagtt
     3721 tccctggccc cacagggaga caccagcacc tagtaaaagt aggagtggga ggagcaagac
     3781 aaaaaccttg caaatatttc gaaggcacct ggtaagaaga aagacattca gccaagaaat
     3841 caaagtaaca aattggtacc agccgctccc tagccaccag aaggttcccg aggctgcctt
     3901 ccgtggtttg agagtcctgt ctctggagta caccctggag gtcatgctag cataacttcg
     3961 tataaagtat cctatacgaa gttatttgcc ttaacccaga aattatcact gttattcttt
     4021 agaatggtgc aaagaataac ttcgtataat gtatgctata cgaagttatg aattcgatat
     4081 caagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     4141 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     4201 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     4261 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     4321 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     4381 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     4441 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt
     4501 tccttggctg ctcgcctatg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     4561 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     4621 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     4681 gcatcgatac cgagcgctgc tcgagagatc tacgggtggc atccctgtga cccctcccca
     4741 gtgcctctcc tggccctgga agttgccact ccagtgccca ccagccttgt cctaataaaa
     4801 ttaagttgca tcattttgtc tgactaggtg tccttctata atattatggg gtggaggggg
     4861 gtggtatgga gcaaggggca agttgggaag acaacctgta gggcctgcgg ggtctattgg
     4921 gaaccaagct ggagtgcagt ggcacaatct tggctcactg caatctccgc ctcctgggtt
     4981 caagcgattc tcctgcctca gcctcccgag ttgttgggat tccaggcatg catgaccagg
     5041 ctcagctaat ttttgttttt ttggtagaga cggggtttca ccatattggc caggctggtc
     5101 tccaactcct aatctcaggt gatctaccca ccttggcctc ccaaattgct gggattacag
     5161 gcgtgaacca ctgctccctt ccctgtcctt ctgattttgt aggtaaccac gtgcggaccg
     5221 agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc
     5281 tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag
     5341 tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc
     5401 atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg
     5461 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     5521 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     5581 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     5641 cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac
     5701 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     5761 tggaacaaca ctcaacccta tctcgggcta ttcttttgat ttataaggga ttttgccgat
     5821 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     5881 aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata
     5941 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     6001 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     6061 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata
     6121 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     6181 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     6241 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     6301 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     6361 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     6421 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     6481 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     6541 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     6601 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     6661 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     6721 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     6781 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     6841 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     6901 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     6961 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     7021 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     7081 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     7141 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     7201 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     7261 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     7321 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     7381 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     7441 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     7501 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     7561 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     7621 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     7681 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     7741 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     7801 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     7861 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     7921 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgt
//
