LOCUS       Exported                6903 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Adeno-associated virus plasmid, Cre-dependent FLEX switch, APEX2NES.
ACCESSION   .
VERSION     .
KEYWORDS    AAV-CAG-DIO-APEX2NES
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6903)
  AUTHORS   Joesch M, Mankus D, Yamagata M, Shahbazi A, Shalek R, Suissa-Peleg 
            A, Meister M, Lichtman JW, Scheirer WJ, Sanes JR
  TITLE     Reconstruction of genetically identified neurons imaged by 
            serial-section electron microscopy.
  JOURNAL   Elife. 2016 Jul 7;5. pii: e15015. doi: 10.7554/eLife.15015.
  PUBMED    27383271
REFERENCE   2  (bases 1 to 6903)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6903
                     /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
     enhancer        156..535
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        537..814
                     /label=chicken beta-actin promoter
     intron          816..1833
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1757..1779
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1841..1860
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    1912..1945
                     /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    1996..2029
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     CDS             complement(2044..2070)
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             complement(2830..2871)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     regulatory      2871..2880
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     protein_bind    complement(2888..2921)
                     /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(2972..3005)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    3030..3618
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3083..3103)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3501..3512)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3650..4126
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3849..3868)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4166..4306
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4166..4295
                     /label=AAV2 ITR
     rep_origin      4381..4836
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4468..4487)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4678..4699
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4853..4872)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4972..4994
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5032..5050)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5118..5222
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5223..6083
                     /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(5441..5460)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6254..6842
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6743..6762
                     /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 cgctcgacat tgattattga ctagttatta
      181 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
      241 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
      301 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
      361 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
      421 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
      481 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtcga
      541 ggtgagcccc acgttctgct tcactctccc catctccccc ccctccccac ccccaatttt
      601 gtatttattt attttttaat tattttgtgc agcgatgggg gcgggggggg ggggggggcg
      661 cgcgccaggc ggggcggggc ggggcgaggg gcggggcggg gcgaggcgga gaggtgcggc
      721 ggcagccaat cagagcggcg cgctccgaaa gtttcctttt atggcgaggc ggcggcggcg
      781 gcggccctat aaaaagcgaa gcgcgcggcg ggcggggagt cgctgcgacg ctgccttcgc
      841 cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta
      901 ctcccacagg tgagcgggcg ggacggccct tctcctccgg gctgtaatta gcgcttggtt
      961 taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc ttgaggggct ccgggagggc
     1021 cctttgtgcg gggggagcgg ctcggggggt gcgtgcgtgt gtgtgtgcgt ggggagcgcc
     1081 gcgtgcggct ccgcgctgcc cggcggctgt gagcgctgcg ggcgcggcgc ggggctttgt
     1141 gcgctccgca gtgtgcgcga ggggagcgcg gccgggggcg gtgccccgcg gtgcgggggg
     1201 ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg tgggggggtg agcagggggt
     1261 gtgggcgcgt cggtcgggct gcaacccccc ctgcaccccc ctccccgagt tgctgagcac
     1321 ggcccggctt cgggtgcggg gctccgtacg gggcgtggcg cggggctcgc cgtgccgggc
     1381 ggggggtggc ggcaggtggg ggtgccgggc ggggcggggc cgcctcgggc cggggagggc
     1441 tcgggggagg ggcgcggcgg cccccggagc gccggcggct gtcgaggcgc ggcgagccgc
     1501 agccattgcc ttttatggta atcgtgcgag agggcgcagg gacttccttt gtcccaaatc
     1561 tgtgcggagc cgaaatctgg gaggcgccgc cgcaccccct ctagcgggcg cggggcgaag
     1621 cggtgcggcg ccggcaggaa ggaaatgggc ggggagggcc ttcgtgcgtc gccgcgccgc
     1681 cgtccccttc tccctctcca gcctcggggc tgtccgcggg gggacggctg ccttcggggg
     1741 ggacggggca gggcggggtt cggcttctgg cgtgtgaccg gcggctctag agcctctgct
     1801 aaccatgttc atgccttctt ctttttccta cagctcctgg gcaacgtgct ggttattgtg
     1861 ctgtctcatc attttggcaa agaattcgga tcctctagag tcgactccgg aataacttcg
     1921 tataggatac tttatacgaa gttatgcaga atggtagctg gattgtagct gctattagca
     1981 atatgaaacc tcttaataac ttcgtatagc atacattata cgaagttatg gcgcgcccta
     2041 gtccagggtc aggcgctcca ggggtggcag ctgcagggcg tcggcaaatc ccagttctga
     2101 cagcttctga tgtgcttctg cgtaatctgc gaaaaaagca tcctcgtcgg ctgcatactt
     2161 atcgaccaga ggtctgaaca cgggatcgga cagcagggct ttgtcgctgg gcagctgcag
     2221 cagcccttcc ttctctccac tcagcagttc agtaaagtaa gaattgtcga aaatcagggg
     2281 gttgcttgtc cagggtccct cgaagcctga cctttcctta tgagcggcgc caatggtgtg
     2341 ccctccagac agtgcgacga tatcctggtc agtcaggccc atagctttcc caaacacatc
     2401 tcgcagatgg tcggagccct ttgtcggatc tggcaggcgc ccctctggag gtggttctgg
     2461 tttgtcctcc cgtcctgggt ggaaaggcac cttggggccc ccagtgacct ccacagcgac
     2521 cactcctgcc agctgataaa aatcggcgta ggacagaatt gggaactctg ctttcagagg
     2581 ttccagcagc cgcacggcga tgtccagccc attgttggca ctatgtgcca gttcagcagg
     2641 gtgcttgatt gttccaaagg gtccgcctgt tttggttccc ttgtcaaagg tgcctgcgct
     2701 gtggaaagcc agtctcagca tcagaggtgc gcacctcttt tctgcgataa accctctcag
     2761 cttcttcttg gctttctcca cggcatcctg gtagtctgcg gacactgttg ggtatgattt
     2821 cccaccggtg gtgctgtcca ggcccagcag ggggttgggg atgggcttgc ccatggtggc
     2881 ggctagcata acttcgtata aagtatccta tacgaagtta tttgccttaa cccagaaatt
     2941 atcactgtta ttctttagaa tggtgcaaag aataacttcg tataatgtat gctatacgaa
     3001 gttatgaatt cgatatcaag cttatcgata atcaacctct ggattacaaa atttgtgaaa
     3061 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     3121 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     3181 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     3241 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     3301 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     3361 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     3421 ggaaatcatc gtcctttcct tggctgctcg cctatgttgc cacctggatt ctgcgcggga
     3481 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     3541 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     3601 tttgggccgc ctccccgcat cgataccgag cgctgctcga gagatctacg ggtggcatcc
     3661 ctgtgacccc tccccagtgc ctctcctggc cctggaagtt gccactccag tgcccaccag
     3721 ccttgtccta ataaaattaa gttgcatcat tttgtctgac taggtgtcct tctataatat
     3781 tatggggtgg aggggggtgg tatggagcaa ggggcaagtt gggaagacaa cctgtagggc
     3841 ctgcggggtc tattgggaac caagctggag tgcagtggca caatcttggc tcactgcaat
     3901 ctccgcctcc tgggttcaag cgattctcct gcctcagcct cccgagttgt tgggattcca
     3961 ggcatgcatg accaggctca gctaattttt gtttttttgg tagagacggg gtttcaccat
     4021 attggccagg ctggtctcca actcctaatc tcaggtgatc tacccacctt ggcctcccaa
     4081 attgctggga ttacaggcgt gaaccactgc tcccttccct gtccttctga ttttgtaggt
     4141 aaccacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct
     4201 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
     4261 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg
     4321 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt
     4381 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     4441 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     4501 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     4561 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc
     4621 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     4681 gactcttgtt ccaaactgga acaacactca accctatctc gggctattct tttgatttat
     4741 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     4801 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct
     4861 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     4921 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     4981 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     5041 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     5101 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     5161 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     5221 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     5281 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     5341 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     5401 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     5461 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     5521 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     5581 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     5641 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     5701 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     5761 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     5821 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     5881 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     5941 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     6001 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     6061 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     6121 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     6181 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     6241 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     6301 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     6361 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     6421 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     6481 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     6541 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     6601 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     6661 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     6721 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     6781 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     6841 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     6901 tgt
//
