LOCUS       pAAV-CAG-FLEX-ArgiNLS-EGFP        5836 bp    DNA     circular UNA 29-APR-2024
DEFINITION  .
ACCESSION   urn.local...4q3-d70cuf4
VERSION     urn.local...4q3-d70cuf4
KEYWORDS    .
SOURCE      
  ORGANISM  .
COMMENT     ##Imported_From-START##
            Path     :: C:\Users\TheGoldenLab\Downloads
            Filename :: addgene-plasmid-28304-sequence-162223.dna
            ##Imported_From-END##
FEATURES             Location/Qualifiers
     repeat_region   56..185
                     /label="AAV2 ITR"
     polyA_signal    347..468
                     /note="<html><body>SV40 polyadenylation
                     signal</body></html>"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(384..403)
                     /Tm=51
                     /Description="SV40 polyA, reverse primer"
                     /label="SV40pA-R"
     primer_bind     438..457
                     /Tm=54
                     /Description="SV40 polyA terminator, reverse primer"
                     /label="EBV-rev"
     misc_feature    511..1099
                     /note="<html><body>woodchuck hepatitis
                     virus&nbsp;posttranscriptional regulatory
                     element</body></html>"
                     /label="WPRE"
     CDS             617..628
                     /codon_start=1
                     /translation="IEGR"
                     /product="<html><body>Factor Xa recognition and cleavage
                     site</body></html>"
                     /transl_table=1
                     /label="Factor Xa site"
     primer_bind     1026..1046
                     /Tm=55
                     /Description="WPRE, reverse primer"
                     /label="WPRE-R"
     protein_bind    complement(1141..1174)
                     /note="<html><body>Cre-mediated recombination occurs in
                     the 8-bp core sequence (GCATACAT).</body></html>"
                     /bound_moiety="<html><body>Cre recombinase</body></html>"
                     /label="loxP"
     protein_bind    complement(1245..1278)
                     /note="<html><body>Cre-mediated recombination occurs in
                     the 8-bp core sequence (GGATACTT). <i>lox2272</i> sites
                     are compatible with each other, but incompatible with
                     <i>loxP</i> or <i>loxN</i> sites.</body></html>"
                     /bound_moiety="<html><body>Cre recombinase</body></html>"
                     /label="lox2272"
     sig_peptide     1304..1375
                     /created_by="TheGoldenLab"
                     /modified_by="ericszelenyi"
                     /label="ArgiNLS"
     CDS             1376..2089
                     /created_by="TheGoldenLab"
                     /label="EGFP"
     primer_bind     complement(1743..1761)
                     /%GC="57.9"
                     /Hairpin_Tm="38.9"
                     /Self_Dimer_Tm="None"
                     /Tm="61.1"
                     /Sequence="TCGATGCCCTTCAGCTCGA"
                     /created_by="primer3"
                     /Transferred_From="GL_2"
                     /Transferred_Similarity="100.00%"
                     /label="GL_2"
     primer_bind     1768..1786
                     /%GC="63.2"
                     /Hairpin_Tm="45.7"
                     /Self_Dimer_Tm="None"
                     /Tm="60.5"
                     /Sequence="GGAGGACGGCAACATCCTG"
                     /created_by="primer3"
                     /Transferred_From="GL_24"
                     /Transferred_Similarity="100.00%"
                     /label="GL_24"
     protein_bind    2115..2148
                     /note="<html><body>Cre-mediated recombination occurs in
                     the 8-bp core sequence (GCATACAT).</body></html>"
                     /bound_moiety="<html><body>Cre recombinase</body></html>"
                     /label="loxP"
     protein_bind    2211..2244
                     /note="<html><body>Cre-mediated recombination occurs in
                     the 8-bp core sequence (GGATACTT). <i>lox2272</i> sites
                     are compatible with each other, but incompatible with
                     <i>loxP</i> or <i>loxN</i> sites.</body></html>"
                     /bound_moiety="<html><body>Cre recombinase</body></html>"
                     /label="lox2272"
     primer_bind     complement(2279..2298)
                     /Tm=57
                     /Description="Rabbit beta-globin intron, for pCAG
                     plasmids, forward primer"
                     /label="pCAG-F"
     primer_bind     complement(2360..2382)
                     /Tm=65
                     /Description="Chimeric intron in CAG promoter, forward
                     primer"
                     /label="pCAGGS-5"
     promoter        complement(2661..2938)
                     /label="chicken β-actin promoter"
     repeat_region   3249..3378
                     /label="AAV2 ITR"
     rep_origin      3450..3905
                     /note="<html><body>f1 bacteriophage origin of replication;
                     arrow indicates direction of (+) strand
                     synthesis</body></html>"
                     /direction="RIGHT"
                     /label="f1 ori"
     primer_bind     complement(3537..3556)
                     /Tm=57
                     /Description="F1 origin, reverse primer"
                     /label="F1ori-R"
     primer_bind     3747..3768
                     /Tm=57
                     /Description="F1 origin, forward primer"
                     /label="F1ori-F"
     promoter        3931..4035
                     /gene="<html><body><i>bla</i></body></html>"
                     /label="AmpR promoter"
     CDS             join(4036..4104,4105..4896)
                     /gene="<html><body><i>bla</i></body></html>"
                     /codon_start=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVG
                     YIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLV
                     EYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTR
                     LDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGP
                     LLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQI
                     AEIGASLIKHW*"
                     /note="<html><body>confers resistance to ampicillin,
                     carbenicillin, and related antibiotics</body></html>"
                     /product="<html><body>β-lactamase</body></html>"
                     /transl_table=1
                     /label="AmpR"
     primer_bind     complement(4254..4273)
                     /Tm=58
                     /Description="Ampicillin resistance gene, reverse primer"
                     /label="Amp-R"
     rep_origin      5067..5655
                     /note="<html><body>high-copy-number ColE1/pMB1/pBR322/pUC
                     origin of replication</body></html>"
                     /direction="RIGHT"
                     /label="ori"
     primer_bind     5556..5575
                     /Tm=56
                     /Description="pBR322 origin, forward primer"
                     /label="pBR322ori-F"
     primer_bind     5809..5826
                     /Tm=58
                     /Description="L4440 vector, forward primer"
                     /label="L4440"
ORIGIN      
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctgcg
       61 cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg
      121 cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg
      181 ttccttgtag ttaatgatta acccgccatg ctacttatct acgtagccat gctctaggaa
      241 gatccttatc gattttacca catttgtaga ggttttactt gctttaaaaa acctcccaca
      301 tctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
      361 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
      421 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgctc
      481 gaagcggccg cccgggtcga cggtatcgat gcggggaggc ggcccaaagg gagatccgac
      541 tcgtctgagg gcgaaggcga agacgcggaa gaggccgcag agccggcagc aggccgcggg
      601 aaggaaggtc cgctggattg agggccgaag ggacgtagca gaaggacgtc ccgcgcagaa
      661 tccaggtggc aacacaggcg agcagccaag gaaaggacga tgatttcccc gacaacacca
      721 cggaattgtc agtgcccaac agccgagccc ctgtccagca gcgggcaagg caggcggcga
      781 tgagttccgc cgtggcaata gggaggggga aagcgaaagt cccggaaagg agctgacagg
      841 tggtggcaat gccccaacca gtgggggttg cgtcagcaaa cacagtgcac accacgccac
      901 gttgcctgac aacgggccac aactcctcat aaagagacag caaccaggat ttatacaagg
      961 aggagaaaat gaaagccata cgggaagcaa tagcatgata caaaggcatt aaagcagcgt
     1021 atccacatag cgtaaaagga gcaacatagt taagaatacc agtcaatctt tcacaaattt
     1081 tgtaatccag aggttgatta tcgataagct tgatatcgaa ttcgaacctc ttcgagggac
     1141 ataacttcgt ataatgtatg ctatacgaag ttatgggtcg atggtgagat ctggactaga
     1201 gggtcgatgg tgatgcttgg atcaaaatag gaagaatccc aatgataact tcgtataaag
     1261 tatcctatac gaagttatac tagaattcgg taccgccacc atgaaaaggg tggctcccca
     1321 gaagcagatg agcatagcaa agaaaaagaa ggtgagaagg cggcggcgcc gtcgagtgag
     1381 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     1441 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     1501 gaccctgaag ttcatttgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     1561 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
     1621 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     1681 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     1741 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     1801 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
     1861 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     1921 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     1981 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     2041 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt gaaactcgag
     2101 gggggatccc tagtataact tcgtatagca tacattatac gaagttatgg gtcgatggtg
     2161 atgcttggca attcgggtcg atggtgaagc attggtcttc ctattttgat ataacttcgt
     2221 ataggatact ttatacgaag ttatggggat ccaattcttt gccaaaatga tgagacagca
     2281 caataaccag cacgttgccc aggagctgta ggaaaaagaa gaaggcatga acatggttag
     2341 cagaggctct agagccgccg gtcacacgcc agaagccgaa ccccgccctg ccccgtcccc
     2401 cccgaaggca gccgtccccc cgcggacagc cccgagccgc tccccccgca caaagggccc
     2461 tcccggagcc cctcaaggct ttcacgcagc cacagaaaag aaacaagccg tcattaaacc
     2521 aagcgctaat tacagcccgg aggagaaggg ccgtcccgcc cgctcacctg tgggagtaac
     2581 gcggtcagtc agagccgggg cgggcggcgc gaggcggcgg cggagcgggg cacggggcga
     2641 aggcagcgcg cagcgactcc cgcccgccgc gcgcttcgct ttttataggg ccgccgccgc
     2701 cgccgcctcg ccataaaagg aaactttcgg agcgcgccgc tctgattggc tgccgccgca
     2761 cctctccgcc tcgccccgcc ccgcccctcg ccccgccccg ccccgcctgg cgcgcgcccc
     2821 cccccccccc ccgcccccat cgctgcacaa aataattaaa aaataaataa atacaaaatt
     2881 gggggtgggg agggggggga gatggggaga gtgaagcaga acgtggggct cacctcgacc
     2941 atggtaatag cgatgactaa tacgtagatg tactgccaag taggaaagtc ccataaggtc
     3001 atgtactggg cataatgcca ggcgggccat ttaccgtcat tgacgtcaat agggggcgta
     3061 cttggcatat gatacacttg atgtactgcc aagtgggcag tttaccgtaa atactccacc
     3121 cattgacgtc aatggaaagt ccctattggc gttactatgg gaacatacgt cattattgac
     3181 gtcaatggta ctcttcctag agcatggcta cgtagataag tagcatggcg ggttaatcat
     3241 taactacaag gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct
     3301 cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt
     3361 gagcgagcga gcgcgcagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
     3421 acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc
     3481 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     3541 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     3601 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     3661 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     3721 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     3781 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     3841 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac
     3901 aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     3961 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     4021 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     4081 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     4141 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     4201 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     4261 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     4321 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     4381 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     4441 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     4501 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     4561 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     4621 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     4681 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     4741 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     4801 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     4861 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     4921 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     4981 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     5041 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     5101 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     5161 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     5221 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     5281 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     5341 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     5401 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     5461 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     5521 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     5581 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     5641 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     5701 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     5761 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     5821 agcgaggaag cggaag
//
