LOCUS       pAAV_Ef1a_DIO_LM        7517 bp DNA     circular SYN 30-JUN-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pAAV Ef1a DIO LMO7
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 7517)
  AUTHORS   Hochgeschwender
  TITLE     Direct Submission
  JOURNAL   Exported Jun 30, 2023 from SnapGene 6.2.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7517
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     repeat_region   12..141
                     /label=AAV2 ITR
     primer_bind     complement(23..34)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     84..95
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        242..1420
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     primer_bind     1368..1388
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     1368..1388
                     /label=C63
     protein_bind    1451..1484
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites."
     protein_bind    1535..1568
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     CDS             complement(1601..2500)
                     /codon_start=1
                     /label=VChR1
                     /translation="MDYPVARSLIVRYPTDLGNGTVCMPRGQCYCEGWLRSRGTSIEKT
                     IAITLQWVVFALSVACLGWYAYQAWRATCGWEEVYVALIEMMKSIIEAFHEFDSPATLW
                     LSSGNGVVWMRYGEWLLTCPVLLIHLSNLTGLKDDYSKRTMGLLVSDVGCIVWGATSAM
                     CTGWTKILFFLISLSYGMYTYFHAAKVYIEAFHTVPKGICRELVRVMAWTFFVAWGMFP
                     VLFLLGTEGFGHISPYGSAIGHSILDLIAKNMWGVLGNYLRVKIHEHILLYGDIRKKQK
                     ITIAGQEMEVETLVAEEED"
     primer_bind     complement(1671..1691)
                     /label=D540
     primer_bind     2392..2415
                     /label=D178
     primer_bind     complement(2473..2493)
                     /label=C492
     CDS             complement(2501..2545)
                     /codon_start=1
                     /label=15 aa Linker
                     /translation="GDPLVQCGGIAGSAT"
     CDS             complement(2546..3046)
                     /codon_start=1
                     /label=eKL9h
                     /translation="FEDFVGDWEQTAAYNLDQVLEQGGVSSVLQTLAVSVTPIQRIVRS
                     GENGLKIDIHVIIPYEGLSADQMAHIEEVFKVVYPVDDHHFKVIMEYGTLVIDGVTPNM
                     LNYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLITPDGSMLFRVTINGVSGWRLFE
                     KISN"
     CDS             complement(3047..3724)
                     /codon_start=1
                     /label=mNeonGreen
                     /translation="VSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYEE
                     LNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFEDG
                     ASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIST
                     FKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQK
                     AFTG"
     primer_bind     3090..3109
                     /label=C454
     primer_bind     complement(3169..3189)
                     /label=C455
     CDS             complement(3725..3775)
                     /codon_start=1
                     /label=Gluc Signal Sequence
                     /translation="MGVKVLFALICIAVAEA"
     protein_bind    complement(3794..3827)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites."
     protein_bind    complement(3878..3911)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    3936..4524
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3989..4009)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     primer_bind     complement(4003..4023)
                     /label=D527
     polyA_signal    4557..4668
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4780..4909
                     /label=AAV2 ITR
     primer_bind     complement(4826..4837)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     4887..4898
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      4995..5450
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5082..5101)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5292..5313
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5467..5486)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5586..5608
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5646..5664)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5732..5836
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5837..6697
                     /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(6055..6074)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6868..7456
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7336..7355
                     /label=C398
     primer_bind     7357..7376
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7406..7425
                     /label=C397
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtaagct ttgcaaagat ggataaagtt
      181 ttaaacagag aggaatcttt gcagctaatg gaccttctag gtcttgaaag gagtgggaat
      241 tggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
      301 gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag
      361 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc
      421 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc
      481 cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat
      541 tacttccacc tggctgcagt acgtgattct tgatcccgag cttcgggttg gaagtgggtg
      601 ggagagttcg aggccttgcg cttaaggagc cccttcgcct cgtgcttgag ttgaggcctg
      661 gcctgggcgc tggggccgcc gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc
      721 tttcgataag tctctagcca tttaaaattt ttgatgacct gctgcgacgc tttttttctg
      781 gcaagatagt cttgtaaatg cgggccaaga tctgcacact ggtatttcgg tttttggggc
      841 cgcgggcggc gacggggccc gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg
      901 agcgcggcca ccgagaatcg gacgggggta gtctcaagct ggccggcctg ctctggtgcc
      961 tggcctcgcg ccgccgtgta tcgccccgcc ctgggcggca aggctggccc ggtcggcacc
     1021 agttgcgtga gcggaaagat ggccgcttcc cggccctgct gcagggagct caaaatggag
     1081 gacgcggcgc tcgggagagc gggcgggtga gtcacccaca caaaggaaaa gggcctttcc
     1141 gtcctcagcc gtcgcttcat gtgactccac ggagtaccgg gcgccgtcca ggcacctcga
     1201 ttagttctcg agcttttgga gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat
     1261 ggagtttccc cacactgagt gggtggagac tgaagttagg ccagcttggc acttgatgta
     1321 attctccttg gaatttgccc tttttgagtt tggatcttgg ttcattctca agcctcagac
     1381 agtggttcaa agtttttttc ttccatttca ggtgtcgtga ggtaccggat cctctagagt
     1441 cgactccgga ataacttcgt ataggatact ttatacgaag ttatgcagaa tggtagctgg
     1501 attgtagctg ctattagcaa tatgaaacct cttaataact tcgtatagca tacattatac
     1561 gaagttatgg cgcgcccaat tccatatgtt aggcggccgc gtcctcctct tcagccacca
     1621 gtgtctcaac ctccatctct tggccggcga tcgtaatttt ctgcttcttt ctgatgtcgc
     1681 catacaacag gatgtgctcg tggattttga cgcgcaggta attccccagc acaccccaca
     1741 tgttctttgc aatcaggtcg aggatggagt gcccaatggc agatccataa gggctgatat
     1801 gaccgaatcc ttccgtgccc agcagaaaca ggactggaaa cattccccaa gccacaaaga
     1861 atgtccaggc catcactcgg accagttctc gacaaatccc ttttggaact gtgtggaatg
     1921 cttcgatata aactttagca gcatgaaaat aggtatacat accatagctc aatgagatga
     1981 ggaagaacag gatcttcgtc caccccgtac acatggcgct ggttgcgccc cacacgatgc
     2041 atcccacatc agaaaccaac aggcccatag tccttttgct ataatcgtcc ttcaggcctg
     2101 tgaggttgct cagatggatc agaaggaccg ggcaggtgag gagccattcg ccataccgca
     2161 tccaaactac gccgttccct gaagacagcc acagtgttgc agggctgtcg aactcgtgga
     2221 aggcctcgat gatgcttttc atcatttcta tcaaggctac gtaaacctcc tcccatccgc
     2281 aggtagctcg ccacgcttga tacgcatacc accccaggca agccactgac aaagcaaaga
     2341 ctacccactg aagagtaatt gctattgtct tttcaatgga agtgccgcga ctccgaagcc
     2401 aaccttcaca gtaacactgt cctcttggca tgcagacggt gccgtttccg aggtcggttg
     2461 ggtagcgaac tatgaggctt ctagcaacag ggtaatccat ggtggcggat ccggcaattc
     2521 caccacactg gactagtggg tcgccgtttg aaatcttctc gaacagccgc cagccgctca
     2581 cgccgttgat ggtcactctg aacagcatag agccatcagg ggtgatcagc cgctcgtcga
     2641 tgatcttatt gccgttccac agggtgcctg tcacggtgat cttcttgcca tcgaacacgg
     2701 cgatgccctc gtaaggccgg ccaaaataat tcagcatgtt tggggtcact ccgtcgatga
     2761 ccagggtgcc atactccatg atcactttga agtggtgatc gtccacgggg tacaccacct
     2821 taaacacctc ctcgatgtgg gccatctgat cggcggacag gccctcgtat gggatgatca
     2881 cgtggatgtc gatcttcagg ccattctcgc cagagcgcac gatcctctgg ataggggtca
     2941 cggacacggc cagtgtctgc agcacggagc tcacgcctcc ctgctccagc acctgatcca
     3001 ggttataggc ggcggtctgc tcccaatcgc ccacgaagtc ctcaaagcct gtgaaggcct
     3061 tctgccactc cttgaagttc agctcggtct tgctgtgctt cagctctgtc ttgcggaaca
     3121 cgtacatggg ctgattcttc agatagttgg cggccattgg cttggcaaag gtatatgtgg
     3181 tcctggctgt agatctgtac cgcttgccgt tgcctgtggt atagctccac ttgaaggtag
     3241 agatgattgt cttatcattg gggtaggtct tcttggatct gcaccagtcg gcggcggtca
     3301 ggctgtttgt catcacgggg ccatcggctg gaaagcctgt gcccttcacc tgggcctcgc
     3361 ccttgatgtg ggagccctcg taggtatatc tgtagttcac tgtcaggctg gcgccgtcct
     3421 cgaactgcat ggtccggtgc acctggtagc cgctgccatc caccatggcg gcctgaaatg
     3481 gagacatgcc gtcagggtag ggcagatact ggtggaagcc atagccgatg tgtggcacca
     3541 ggatccaagg agaaaactgc aggtcgccct tggtggactt cagattcagc tcctcgtagc
     3601 catcattagg gttgcctgtg ccctggccca ccatgtcaaa atccaccccg ttgatgctcc
     3661 caaaaatgtg cagctcgtga gtagcgggca gactagccat attatcctcc tctcctttag
     3721 acacggcctc ggccacagcg atgcagatca gggcaaacag aactttgact cccatggtac
     3781 cgtcgacgct agcataactt cgtataaagt atcctatacg aagttatttg ccttaaccca
     3841 gaaattatca ctgttattct ttagaatggt gcaaagaata acttcgtata atgtatgcta
     3901 tacgaagtta tgaattcgat atcaagctta tcgataatca acctctggat tacaaaattt
     3961 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     4021 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     4081 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     4141 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     4201 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     4261 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     4321 tgtcggggaa atcatcgtcc tttccttggc tgctcgccta tgttgccacc tggattctgc
     4381 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     4441 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     4501 tctccctttg ggccgcctcc ccgcatcgat accgagcgct gctcgagaga tctcgactgt
     4561 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     4621 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     4681 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     4741 agacaatagc aggcatgcac gtgcggaccg agcggccgca ggaaccccta gtgatggagt
     4801 tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc
     4861 gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg
     4921 ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc
     4981 aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     5041 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     5101 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     5161 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg
     5221 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     5281 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcgggcta
     5341 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     5401 ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac
     5461 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     5521 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     5581 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     5641 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     5701 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     5761 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     5821 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     5881 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     5941 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     6001 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     6061 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     6121 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     6181 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     6241 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     6301 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     6361 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     6421 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     6481 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     6541 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     6601 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     6661 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     6721 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     6781 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     6841 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     6901 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     6961 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     7021 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     7081 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     7141 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     7201 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     7261 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     7321 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     7381 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     7441 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     7501 gccttttgct cacatgt
//
