LOCUS       Exported                7029 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  AAV expression of GFP-Cre from CamKII promoter.
ACCESSION   .
VERSION     .
KEYWORDS    pENN.AAV.CamKII.HI.GFP-Cre.WPRE.SV40
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7029)
  TITLE     AAV plasmids
REFERENCE   2  (bases 1 to 7029)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7029
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   111..240
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1783..1792
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1789..2505
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             1789..2505
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1834..1855)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2095..2114)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2442..2463
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2512..2541
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2548..2568
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2569..3597
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="SNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKMLL
                     SVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGLP
                     RPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAFL
                     GIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVER
                     WISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQR
                     YLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLED
                     GD"
     misc_feature    3618..4206
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3671..3691)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4089..4100)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4239..4373
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4276..4295)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4330..4349
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   4402..4542
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4402..4531
                     /label=AAV2 ITR
     rep_origin      4617..5072
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4704..4723)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4914..4935
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5089..5108)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5208..5230
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5268..5286)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5354..5458
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5459..6319
                     /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(5677..5696)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6490..49
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6979..6998
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
       61 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt cctgcaggca
      121 gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt ggtcgcccgg
      181 cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct
      241 gcggccgcac gcgtactagg tagccatgct ctaggaagat ctaacattat ggccttaggt
      301 cacttcatct ccatggggtt cttcttctga ttttctagaa aatgagatgg gggtgcagag
      361 agcttcctca gtgacctgcc cagggtcaca tcagaaatgt cagagctaga acttgaactc
      421 agattactaa tcttaaattc catgccttgg gggcatgcaa gtacgatata cagaaggagt
      481 gaactcatta gggcagatga ccaatgagtt taggaaagaa gagtccaggg cagggtacat
      541 ctacaccacc cgcccagccc tgggtgagtc cagccacgtt cacctcatta tagttgcctc
      601 tctccagtcc taccttgacg ggaagcacaa gcagaaactg ggacaggagc cccaggagac
      661 caaatcttca tggtccctct gggaggatgg gtggggagag ctgtggcaga ggcctcagga
      721 ggggccctgc tgctcagtgg tgacagatag gggtgagaaa gcagacagag tcattccgtc
      781 agcattctgg gtctgtttgg tacttcttct cacgctaagg tggcggtgtg atatgcacaa
      841 tggctaaaaa gcagggagag ctggaaagaa acaaggacag agacagaggc caagtcaacc
      901 agaccaattc ccagaggaag caaagaaacc attacagaga ctacaagggg gaagggaagg
      961 agagatgaat tagcttcccc tgtaaacctt agaacccagc tgttgccagg gcaacggggc
     1021 aatacctgtc tcttcagagg agatgaagtt gccagggtaa ctacatcctg tctttctcaa
     1081 ggaccatccc agaatgtggc acccactagc cgttaccata gcaactgcct ctttgcccca
     1141 cttaatccca tcccgtctgt taaaagggcc ctatagttgg aggtggggga ggtaggaaga
     1201 gcgatgatca cttgtggact aagtttgttc gcatcccctt ctccaacccc ctcagtacat
     1261 caccctgggg gaacagggtc cacttgctcc tgggcccaca cagtcctgca gtattgtgta
     1321 tataaggcca gggcaaagag gagcaggttt taaagtgaaa ggcaggcagg tgttggggag
     1381 gcagttaccg gggcaacggg aacagggcgt ttcggaggtg gttgccatgg ggacctggat
     1441 gctgacgaag gctcgcggga ccgatccagc ctccgtaccg gttcgaacag gtaagcgccc
     1501 ctaaaatccc tttgggcaca atgtgtcctg aggggagagg cagcgacctg tagatgggac
     1561 gggggcacta accctcaggt ttggggcttc tgaatgtgag tatcgccatg taagcccagt
     1621 atttggccaa tctcagaaag ctcctggtcc ctggagggat ggagagagaa aaacaaacag
     1681 ctcctggagc agggagagtg ctggcctctt gctctccggc tccctctgtt gccctctggt
     1741 ttctccccag gttcgaatct agaggatccc cgggtaccgg tcgccaccat ggtgagcaag
     1801 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     1861 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
     1921 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     1981 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
     2041 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     2101 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     2161 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     2221 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg
     2281 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     2341 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     2401 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     2461 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtccgg agagcaaaag
     2521 ctgatttctg aggaggatct gggaggaccc aagaagaaga ggaaggtgtc caatttactg
     2581 accgtacacc aaaatttgcc tgcattaccg gtcgatgcaa cgagtgatga ggttcgcaag
     2641 aacctgatgg acatgttcag ggatcgccag gcgttttctg agcatacctg gaaaatgctt
     2701 ctgtccgttt gccggtcgtg ggcggcatgg tgcaagttga ataaccggaa atggtttccc
     2761 gcagaacctg aagatgttcg cgattatctt ctatatcttc aggcgcgcgg tctggcagta
     2821 aaaactatcc agcaacattt gggccagcta aacatgcttc atcgtcggtc cgggctgcca
     2881 cgaccaagtg acagcaatgc tgtttcactg gttatgcggc ggatccgaaa agaaaacgtt
     2941 gatgccggtg aacgtgcaaa acaggctcta gcgttcgaac gcactgattt cgaccaggtt
     3001 cgttcactca tggaaaatag cgatcgctgc caggatatac gtaatctggc atttctgggg
     3061 attgcttata acaccctgtt acgtatagcc gaaattgcca ggatcagggt taaagatatc
     3121 tcacgtactg acggtgggag aatgttaatc catattggca gaacgaaaac gctggttagc
     3181 accgcaggtg tagagaaggc acttagcctg ggggtaacta aactggtcga gcgatggatt
     3241 tccgtctctg gtgtagctga tgatccgaat aactacctgt tttgccgggt cagaaaaaat
     3301 ggtgttgccg cgccatctgc caccagccag ctatcaactc gcgccctgga agggattttt
     3361 gaagcaactc atcgattgat ttacggcgct aaggatgact ctggtcagag atacctggcc
     3421 tggtctggac acagtgcccg tgtcggagcc gcgcgagata tggcccgcgc tggagtttca
     3481 ataccggaga tcatgcaagc tggtggctgg accaatgtaa atattgtcat gaactatatc
     3541 cgtaacctgg atagtgaaac aggggcaatg gtgcgcctgc tggaagatgg cgattagctc
     3601 gaatcaagct tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     3661 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     3721 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     3781 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     3841 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     3901 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     3961 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt
     4021 cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct
     4081 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     4141 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     4201 ccccgcatcg ataccgtcga tcgagaaatc gatcgcccaa cttgtttatt gcagcttata
     4261 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     4321 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg atctccggac
     4381 acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc
     4441 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc
     4501 gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt
     4561 ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc
     4621 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     4681 acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     4741 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     4801 tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc
     4861 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     4921 cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg atttataagg
     4981 gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     5041 gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc
     5101 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
     5161 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     5221 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg
     5281 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     5341 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     5401 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     5461 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     5521 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     5581 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     5641 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     5701 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     5761 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     5821 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     5881 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     5941 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     6001 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     6061 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     6121 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     6181 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     6241 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     6301 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     6361 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     6421 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     6481 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     6541 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     6601 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     6661 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     6721 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     6781 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     6841 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     6901 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     6961 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     7021 cctctgact
//
