LOCUS       Exported                6476 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  AAV expression of Cre-GFP fusion.
ACCESSION   .
VERSION     .
KEYWORDS    AAV-Cre-GFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6476)
  AUTHORS   Kennedy PJ, Feng J, Robison AJ, Maze I, Badimon A, Mouzon E, 
            Chaudhury D, Damez-Werno DM, Haggarty SJ, Han MH, Bassel-Duby R, 
            Olson EN, Nestler EJ
  TITLE     Class I HDAC inhibition blocks cocaine-induced plasticity by 
            targeted changes in histone methylation.
  JOURNAL   Nat Neurosci. 2013 Apr;16(4):434-40. doi: 10.1038/nn.3354. Epub 2013
            Mar 10.
  PUBMED    23475113
REFERENCE   2  (bases 1 to 6476)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6476
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             1..717
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             1..717
                     /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(46..67)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(307..326)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     654..675
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             772..1803
                     /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="MANLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     polyA_signal    1880..2356
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2079..2098)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2396..2536
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2396..2525
                     /label=AAV2 ITR
     rep_origin      2611..3066
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2698..2717)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2908..2929
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3083..3102)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3202..3224
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3262..3280)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3348..3452
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3453..4313
                     /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(3671..3690)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4484..5072
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4973..4992
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   5134..5263
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        5339..5642
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5643..5846
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5797..5817
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5843..5867
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      6471..4
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
ORIGIN
        1 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
       61 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
      121 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
      181 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
      241 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
      301 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
      361 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
      421 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
      481 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
      541 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
      601 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
      661 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtcc
      721 ggtacagctc tcgacggaga aagctcaggc tctggctcag agtctgactc catggccaat
      781 ttactgaccg tacaccaaaa tttgcctgca ttaccggtcg atgcaacgag tgatgaggtt
      841 cgcaagaacc tgatggacat gttcagggat cgccaggcgt tttctgagca tacctggaaa
      901 atgcttctgt ccgtttgccg gtcgtgggcg gcatggtgca agttgaataa ccggaaatgg
      961 tttcccgcag aacctgaaga tgttcgcgat tatcttctat atcttcaggc gcgcggtctg
     1021 gcagtaaaaa ctatccagca acatttgggc cagctaaaca tgcttcatcg tcggtccggg
     1081 ctgccacgac caagtgacag caatgctgtt tcactggtta tgcggcggat ccgaaaagaa
     1141 aacgttgatg ccggtgaacg tgcaaaacag gctctagcgt tcgaacgcac tgatttcgac
     1201 caggttcgtt cactcatgga aaatagcgat cgctgccagg atatacgtaa tctggcattt
     1261 ctggggattg cttataacac cctgttacgt atagccgaaa ttgccaggat cagggttaaa
     1321 gatatctcac gtactgacgg tgggagaatg ttaatccata ttggcagaac gaaaacgctg
     1381 gttagcaccg caggtgtaga gaaggcactt agcctggggg taactaaact ggtcgagcga
     1441 tggatttccg tctctggtgt agctgatgat ccgaataact acctgttttg ccgggtcaga
     1501 aaaaatggtg ttgccgcgcc atctgccacc agccagctat caactcgcgc cctggaaggg
     1561 atttttgaag caactcatcg attgatttac ggcgctaagg atgactctgg tcagagatac
     1621 ctggcctggt ctggacacag tgcccgtgtc ggagccgcgc gagatatggc ccgcgctgga
     1681 gtttcaatac cggagatcat gcaagctggt ggctggacca atgtaaatat tgtcatgaac
     1741 tatatccgta acctggatag tgaaacaggg gcaatggtgc gcctgctgga agatggcgat
     1801 tagccattaa cgcgtaaatg attgctctag agtcgacctg cagaagcttg cctcgagcag
     1861 cgctgctcga gagatctacg ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc
     1921 cctggaagtt gccactccag tgcccaccag ccttgtccta ataaaattaa gttgcatcat
     1981 tttgtctgac taggtgtcct tctataatat tatggggtgg aggggggtgg tatggagcaa
     2041 ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc tattgggaac caagctggag
     2101 tgcagtggca caatcttggc tcactgcaat ctccgcctcc tgggttcaag cgattctcct
     2161 gcctcagcct cccgagttgt tgggattcca ggcatgcatg accaggctca gctaattttt
     2221 gtttttttgg tagagacggg gtttcaccat attggccagg ctggtctcca actcctaatc
     2281 tcaggtgatc tacccacctt ggcctcccaa attgctggga ttacaggcgt gaaccactgc
     2341 tcccttccct gtccttctga ttttgtaggt aaccacgtgc ggaccgagcg gccgcaggaa
     2401 cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg
     2461 cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg
     2521 cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     2581 tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg
     2641 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc
     2701 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     2761 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     2821 ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     2881 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     2941 actctatctc gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt
     3001 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta
     3061 caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     3121 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     3181 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     3241 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     3301 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     3361 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     3421 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     3481 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     3541 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     3601 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     3661 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     3721 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     3781 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     3841 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     3901 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     3961 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     4021 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     4081 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     4141 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     4201 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     4261 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     4321 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     4381 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     4441 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     4501 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     4561 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     4621 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     4681 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     4741 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     4801 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     4861 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     4921 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     4981 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     5041 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     5101 tcctggcctt ttgctggcct tttgctcaca tgtcctgcag gcagctgcgc gctcgctcgc
     5161 tcactgaggc cgcccgggcg tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga
     5221 gcgcgcagag agggagtggc caactccatc actaggggtt cctgcggccg cacgcgtcta
     5281 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
     5341 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
     5401 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
     5461 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
     5521 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
     5581 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
     5641 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
     5701 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
     5761 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
     5821 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagatcgcc tggagacgcc
     5881 atccacgctg ttttgacctc catagaagac accgggaccg atccagcctc cgcggattcg
     5941 aatcccggcc gggaacggtg cattggaacg cggattcccc gtgccaagag tgacgtaagt
     6001 accgcctata gagtctatag gcccacaaaa aatgctttct tcttttaata tacttttttg
     6061 tttatcttat ttctaatact ttccctaatc tctttctttc agggcaataa tgatacaatg
     6121 tatcatgcct ctttgcacca ttctaaagaa taacagtgat aatttctggg ttaaggcaat
     6181 agcaatattt ctgcatataa atatttctgc atataaattg taactgatgt aagaggtttc
     6241 atattgctaa tagcagctac aatccagcta ccattctgct tttattttat ggttgggata
     6301 aggctggatt attctgagtc caagctaggc ccttttgcta atcatgttca tacctcttat
     6361 cttcctccca cagctcctgg gcaacgtgct ggtctgtgtg ctggcccatc actttggcaa
     6421 agaattggga ttcgaacatc gattgaattc cccggggatc tgcaccggtc gccacc
//
