LOCUS       Exported                6087 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Can be used to express Cre recombinase and simultaneously the red 
            fluorescent protein dTomato. Can also be used to create 
            adeno-associated virus for delivery of these genes..
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-hSyn-Cre-P2A-dTomato
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6087)
  TITLE     Cre-expressing AAV Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6087)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6087
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        183..630
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      654..663
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             666..686
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1722..1778
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             1779..2480
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLYGMDELYK"
     primer_bind     complement(1912..1930)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2180..2199
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2374..2397
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     misc_feature    2499..3087
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2552..2572)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2970..2981)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3121..3232
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3350..3490
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3350..3479
                     /label=AAV2 ITR
     rep_origin      3565..4020
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3652..3671)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3862..3883
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4037..4056)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4156..4178
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4216..4234)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4302..4406
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4407..5267
                     /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(4625..4644)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5438..6026
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5927..5946
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtttaatt aagtgtctag actgcagagg gccctgcgta
      181 tgagtgcaag tgggttttag gaccaggatg aggcggggtg ggggtgccta cctgacgacc
      241 gaccccgacc cactggacaa gcacccaacc cccattcccc aaattgcgca tcccctatca
      301 gagaggggga ggggaaacag gatgcggcga ggcgcgtgcg cactgccagc ttcagcaccg
      361 cggacagtgc cttcgccccc gcctggcggc gcgcgccacc gccgcctcag cactgaaggc
      421 gcgctgacgt cactcgccgg tcccccgcaa actccccttc ccggccacct tggtcgcgtc
      481 cgcgccgccg ccggcccagc cggaccgcac cacgcgaggc gcgagatagg ggggcacggg
      541 cgcgaccatc tgcgctgcgg cgccggcgac tcagcgctgc ctcagtctgc ggtgggcagc
      601 ggaggagtcg tgtcgtgcct gagagcgcag tcgagaaacc ggctagagga tccgccacca
      661 tggtgcccaa gaagaagagg aaagtctcca acctgctgac tgtgcaccaa aacctgcctg
      721 ccctccctgt ggatgccacc tctgatgaag tcaggaagaa cctgatggac atgttcaggg
      781 acaggcaggc cttctctgaa cacacctgga agatgctcct gtctgtgtgc agatcctggg
      841 ctgcctggtg caagctgaac aacaggaaat ggttccctgc tgaacctgag gatgtgaggg
      901 actacctcct gtacctgcaa gccagaggcc tggctgtgaa gaccatccaa cagcacctgg
      961 gccagctcaa catgctgcac aggagatctg gcctgcctcg cccttctgac tccaatgctg
     1021 tgtccctggt gatgaggaga atcagaaagg agaatgtgga tgctggggag agagccaagc
     1081 aggccctggc ctttgaacgc actgactttg accaagtcag atccctgatg gagaactctg
     1141 acagatgcca ggacatcagg aacctggcct tcctgggcat tgcctacaac accctgctgc
     1201 gcattgccga aattgccaga atcagagtga aggacatctc ccgcaccgat ggtgggagaa
     1261 tgctgatcca cattggcagg accaagaccc tggtgtccac agctggtgtg gagaaggccc
     1321 tgtccctggg ggttaccaag ctggtggaga gatggatctc tgtgtctggt gtggctgatg
     1381 accccaacaa ctacctgttc tgccgggtca gaaagaatgg tgtggctgcc ccttctgcca
     1441 cctcccaact gtccacccgg gccctggaag ggatctttga ggccacccac cgcctgatct
     1501 atggtgccaa ggatgactct gggcagagat acctggcctg gtctggccac tctgccagag
     1561 tgggtgctgc cagggacatg gccagggctg gtgtgtccat ccctgaaatc atgcaggctg
     1621 gtggctggac caatgtgaac atagtgatga actacatcag aaacctggac tctgagactg
     1681 gggccatggt gaggctgctc gaggatgggg acggaagcgg agccactaac ttctccctgt
     1741 tgaaacaagc aggggatgtc gaagagaatc ccgggccaat ggtgagcaag ggcgaggagg
     1801 tcatcaaaga gttcatgcgc ttcaaggtgc gcatggaggg ctccatgaac ggccacgagt
     1861 tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga
     1921 aggtgaccaa gggcggcccc ctgcccttcg cctgggacat cctgtccccc cagttcatgt
     1981 acggctccaa ggcgtacgtg aagcaccccg ccgacatccc cgattacaag aagctgtcct
     2041 tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggt ctggtgaccg
     2101 tgacccagga ctcctccctg caggacggca cgctgatcta caaggtgaag atgcgcggca
     2161 ccaacttccc ccccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcca
     2221 ccgagcgcct gtacccccgc gacggcgtgc tgaagggcga gatccaccag gccctgaagc
     2281 tgaaggacgg cggccactac ctggtggagt tcaagaccat ctacatggcc aagaagcccg
     2341 tgcaactgcc cggctactac tacgtggaca ccaagctgga catcacctcc cacaacgagg
     2401 actacaccat cgtggaacag tacgagcgct ccgagggccg ccaccacctg ttcctgtacg
     2461 gcatggacga gctgtacaag tagtaaaagc ttatcgataa tcaacctctg gattacaaaa
     2521 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
     2581 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
     2641 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg
     2701 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct
     2761 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg
     2821 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     2881 tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctatgttgcc acctggattc
     2941 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc
     3001 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc
     3061 ggatctccct ttgggccgcc tccccgcatc gataccgagc gctgctcgag agatcttcga
     3121 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     3181 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     3241 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     3301 gggaagacaa tagcaggcat gagatctcac gtgcggaccg agcggccgca ggaaccccta
     3361 gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca
     3421 aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc
     3481 tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     3541 cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     3601 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cttagcgccc gctcctttcg
     3661 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     3721 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     3781 tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     3841 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaactcta
     3901 tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggtctat tggttaaaaa
     3961 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt
     4021 tatggtgcac tctcagtaca atctgctctg atgccgcata gttacgccag ccccgacacc
     4081 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     4141 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     4201 gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     4261 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     4321 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     4381 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     4441 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     4501 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     4561 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     4621 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     4681 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     4741 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     4801 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     4861 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     4921 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     4981 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     5041 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     5101 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     5161 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     5221 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     5281 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     5341 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     5401 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     5461 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     5521 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     5581 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     5641 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     5701 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     5761 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     5821 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     5881 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     5941 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     6001 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     6061 ccttttgctg gccttttgct cacatgt
//
