LOCUS       Exported                6878 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Can be used to generate AAV virus that will express Cre in neurons 
            from the synapsin promoter.
ACCESSION   .
VERSION     .
KEYWORDS    AAV pmSyn1-EBFP-Cre
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6878)
  AUTHORS   Madisen L, Garner AR, Shimaoka D, Chuong AS, Klapoetke NC, Li L, van
            der Bourg A, Niino Y, Egolf L, Monetti C, Gu H, Mills M, Cheng A, 
            Tasic B, Nguyen TN, Sunkin SM, Benucci A, Nagy A, Miyawaki A, 
            Helmchen F, Empson RM, Knopfel T, Boyden ES, Reid RC, Carandini M, 
            Zeng H
  TITLE     Transgenic mice for intersectional targeting of neural sensors and 
            effectors with high specificity and performance.
  JOURNAL   Neuron. 2015 Mar 4;85(5):942-58. doi: 10.1016/j.neuron.2015.02.022.
  PUBMED    25741722
REFERENCE   2  (bases 1 to 6878)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6878
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1590..1719
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(1781..2369)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1861..1880)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2540..3400)
                     /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     3163..3182
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3401..3505)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3573..3591
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3629..3651)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3751..3770
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(3787..4242)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3924..3945)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4136..4155
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   4317..4457
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4328..4457
                     /label=AAV2 ITR
     polyA_signal    4497..4973
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     4755..4774
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     CDS             complement(5047..6078)
                     /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"
     CDS             complement(6091..6807)
                     /codon_start=1
                     /product="enhanced blue variant of GFP (Ai et al., 2007)"
                     /label=EBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIK
                     VNFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLL
                     EFRTAAGITLGMDELYK"
     primer_bind     6741..6762
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      6804..6813
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
ORIGIN
        1 gggctcagga gcacggcggt gctctctggc acgacgcgac tcctctgctg cccgctgcag
       61 actgaggcag cgccgaggct ctgccgccgc agagcgtatg gtcgcgcccg cacctcccta
      121 tcttgcacct ctcgtggtgc ggtccgactg gactcacggc agcatggacg cgaccaaggc
      181 ggcgaggaag gggagtttgg ggagggctag tgagtgacgt cagtcagcct ttggtgctgg
      241 ggaggtggtg gtgcgcgccg gcagcggagg cgaaggctct gaccgcggtg ctgaatccga
      301 cagtgcgcat gcgccgcgcc gcaacctctt cccctccccc tctgatgggg gcgggggatg
      361 cgcaatttgg ggagtggggg tggggttctt gtcctgaggg tccggacctg tagtaacgaa
      421 actccctccc acgatcctac cctgtttacc aacctcttcc tattccttag gctaacttgc
      481 acttacacac tcttgccctt cccagatact tgggtttcca gtgttgattt tcacctttga
      541 aggcattttg gggatagaag cacactcaga ggggaacata aatgtagaaa ttctccttcc
      601 tgcagtgttc ccggggagac atctcctata acaactctgt cgggcaaaga aaagagcacg
      661 ggggtggcgt aaacatcacc agacaggcgg aggtggacac ttgggtagtt aacatatcct
      721 tgcccttggt ggtgagaatc tgaaagaaca catggaataa cacaaataga aaaggaagga
      781 aagacagaat atatatgtac atcacagaac acacaggcat ccaggtatat atgtagacac
      841 agaagctcag ggtaacacac atgagcatat tgagcacaca ttacacattc agataacact
      901 ttgactcaca cccagagaat ctcacacaag tgcacacatc taggaaaaac acagactccc
      961 agatgagcaa ggaaactctg ggtatgcaat gcagaagcat ttcctgcagc gctagctctg
     1021 atgtgttttc agcctcctcc acatgcacat atttagtcta catggcctca ttcctctgct
     1081 ccggacatag cccaaaccca gtgtcattag tgttcatgga tgtcttcctc tttgcagaca
     1141 ggaagccaca taatactttc atgtggaaaa ataaagtaat tttggtggtc tgaataagaa
     1201 ggctcatata tttgaatgct tagtcaccag ggactggtgc tgtttggaaa gattaaaagg
     1261 attagaatgt gtggccttgt tggagaaagc acgtcactgg ggtgggtttt gggggttcaa
     1321 aagtccatgc caagcccaga gccagccagc cagccagcca gccagccagt cagtctgtct
     1381 gtctgtctgt ctgtctgtct gtctgtgtct gttgtctgtg gatcagcata tagctctcag
     1441 ctatggctcc agcatcaagc ctgtgtctgc cacaacactc ctgattatgg taataattta
     1501 ctcatctctg aaactgtaag aaagctccca attaaatgtt tttttttttt taatgagaac
     1561 tgccttggtg atggtacgcg tgcggccgca ggaaccccta gtgatggagt tggccactcc
     1621 ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg
     1681 cggcctcagt gagcgagcga gcgcgcagct gcctgcagga catgtgagca aaaggccagc
     1741 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     1801 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     1861 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     1921 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     1981 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     2041 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     2101 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     2161 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     2221 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     2281 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     2341 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     2401 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     2461 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     2521 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     2581 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     2641 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     2701 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     2761 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     2821 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     2881 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     2941 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     3001 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     3061 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     3121 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     3181 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     3241 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     3301 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     3361 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     3421 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     3481 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     3541 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     3601 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     3661 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     3721 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     3781 accataaaat tgtaaacgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     3841 gctcattttt taaccaatag accgaaatcg gcaaaatccc ttataaatca aaagaatagc
     3901 ccgagataga gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     3961 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     4021 cacccaaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     4081 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     4141 agaaagcgaa aggagcgggc gctaaggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     4201 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtactatggt tgctttgacg
     4261 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgcccctg
     4321 caggcagctg cgcgctcgct cgctcactga ggccgcccgg gcaaagcccg ggcgtcgggc
     4381 gacctttggt cgcccggcct cagtgagcga gcgagcgcgc agagagggag tggccaactc
     4441 catcactagg ggttcctgcg gccgctcggt ccgcacgtgg ttacctacaa aatcagaagg
     4501 acagggaagg gagcagtggt tcacgcctgt aatcccagca atttgggagg ccaaggtggg
     4561 tagatcacct gagattagga gttggagacc agcctggcca atatggtgaa accccgtctc
     4621 taccaaaaaa acaaaaatta gctgagcctg gtcatgcatg cctggaatcc caacaactcg
     4681 ggaggctgag gcaggagaat cgcttgaacc caggaggcgg agattgcagt gagccaagat
     4741 tgtgccactg cactccagct tggttcccaa tagaccccgc aggccctaca ggttgtcttc
     4801 ccaacttgcc ccttgctcca taccaccccc ctccacccca taatattata gaaggacacc
     4861 tagtcagaca aaatgatgca acttaatttt attaggacaa ggctggtggg cactggagtg
     4921 gcaacttcca gggccaggag aggcactggg gaggggtcac agggatgcca cccgtagatc
     4981 tctcgagcag cgctgctcga ggcaagcttc tgcaggtcga ctctagagga tcccccgcgt
     5041 taatggctaa tcgccatctt ccagcaggcg caccattgcc cctgtttcac tatccaggtt
     5101 acggatatag ttcatgacaa tatttacatt ggtccagcca ccagcttgca tgatctccgg
     5161 tattgaaact ccagcgcggg ccatatctcg cgcggctccg acacgggcac tgtgtccaga
     5221 ccaggccagg tatctctgac cagagtcatc cttagcgccg taaatcaatc gatgagttgc
     5281 ttcaaaaatc ccttccaggg cgcgagttga tagctggctg gtggcagatg gcgcggcaac
     5341 accatttttt ctgacccggc aaaacaggta gttattcgga tcatcagcta caccagagac
     5401 ggaaatccat cgctcgacca gtttagttac ccccaggcta agtgccttct ctacacctgc
     5461 ggtgctaacc agcgttttcg ttctgccaat atggattaac attctcccac cgtcagtacg
     5521 tgagatatct ttaaccctga tcctggcaat ttcggctata cgtaacaggg tgttataagc
     5581 aatccccaga aatgccagat tacgtatatc ctggcagcga tcgctatttt ccatgagtga
     5641 acgaacctgg tcgaaatcag tgcgttcgaa cgctagagcc tgttttgcac gttcaccggc
     5701 atcaacgttt tcttttcgga tccgccgcat aaccagtgaa acagcattgc tgtcacttgg
     5761 tcgtggcagc ccggaccgac gatgaagcat gtttagctgg cccaaatgtt gctggatagt
     5821 ttttactgcc agaccgcgcg cctgaagata tagaagataa tcgcgaacat cttcaggttc
     5881 tgcgggaaac catttccggt tattcaactt gcaccatgcc gcccacgacc ggcaaacgga
     5941 cagaagcatt ttccaggtat gctcagaaaa cgcctggcga tccctgaaca tgtccatcag
     6001 gttcttgcga acctcatcac tcgttgcatc gaccggtaat gcaggcaaat tttggtgtac
     6061 ggtcagtaaa ttggccatgg aaccggaacc cttgtacagc tcgtccatgc cgagagtgat
     6121 cccggcggcg gtgcggaact ccagcaggac catgtgatcg cgcttctcgt tggggtcttt
     6181 gctcagcacg gactgggtgc tcaggtagtg gctgtcgggc agcagcacgg ggccgtcgcc
     6241 gatgggggtg ttctgctggt agtggtcggc gagctgcacg ctgccgtcct ccacgttgtg
     6301 gcggatcttg aagttcacct tgatgccgtt cttctgcttg acggccatga tatagatgtt
     6361 gtggctgttg aagttgtact ccagcttgtg ccccaggatg ttgccgtcct ccttgaagtc
     6421 gacgcccttc agctcgatgc ggttcactag ggtgtcgccc tcgaacttca cctcggcgcg
     6481 ggtcttgtag gtgccgtcgt ccttgaagaa gatggtgcgc tcctggacgt agccttcggg
     6541 catggcggac ttgaagaagt cgtgctgctt catgtggtcg gggtagcggg cgaagcactg
     6601 cacgccgtgg ctcagggtgg tcacgagggt gggccagggc acgggcagct tgccggtggt
     6661 gcagatgaac ttcagggtca gcttgccgtt ggtggcatcg ccctcgccct cgcccctcac
     6721 gctgaacttg tggccgttta cgtcgccgtc cagctcgacc aggatgggca ccaccccggt
     6781 gaacagctcc tcgcccttgc tcaccatggt ggcgaccggt gcagatcggg gaattccaag
     6841 gggcagtggg tctgtgggcc gcgggggcgg agcgcgag
//
