LOCUS       Exported                7181 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  pAAV vector to express PBG (chimeric Glycoprotein) in a 
            Cre-dependent manner.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-Ef1a-DIO-PBG-WPRE-hGH
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7181)
  AUTHORS   Kim EJ, Juavinett AL, Kyubwa EM, Jacobs MW, Callaway EM
  TITLE     Three Types of Cortical Layer 5 Neurons That Differ in Brain-wide 
            Connectivity and Function.
  JOURNAL   Neuron. 2015 Dec 16;88(6):1253-67. doi: 
            10.1016/j.neuron.2015.11.002. Epub 2015 Dec 6.
  PUBMED    26671462
REFERENCE   2  (bases 1 to 7181)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7181
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     promoter        242..1419
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          473..1410
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1367..1387
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    1450..1483
                     /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 (Lee and 
                     Saito, 1988)."
     protein_bind    1534..1567
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     regulatory      3147..3156
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     protein_bind    complement(3166..3199)
                     /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 (Lee and 
                     Saito, 1988)."
     protein_bind    complement(3250..3283)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    3308..3896
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3361..3381)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3779..3790)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3928..4404
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4127..4146)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4444..4584
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4444..4573
                     /label=AAV2 ITR
     rep_origin      4659..5114
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4746..4765)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4956..4977
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5131..5150)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5250..5272
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5310..5328)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5396..5500
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5501..6361
                     /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(5719..5738)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6532..7120
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7021..7040
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
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 tacttccact ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      601 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      661 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      721 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      781 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      841 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      901 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      961 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
     1021 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
     1081 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
     1141 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
     1201 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1261 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1321 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1381 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gtaccggatc ctctagagtc
     1441 gactccggaa taacttcgta taggatactt tatacgaagt tatgcagaat ggtagctgga
     1501 ttgtagctgc tattagcaat atgaaacctc ttaataactt cgtatagcat acattatacg
     1561 aagttatggc gcgccttaca gtctggtctc acccccactc ttgtgtgatt cccatgaaga
     1621 tatgatcttc ccgctttggg gagtgactga cacctccctc cctgtccctc tgagattgtg
     1681 ttgcgtaggt tctgatcgat tgactcttct ccagcatgtc atcaggaaaa ttatcaacat
     1741 caaggcagtc agggcccctg cactcagtaa tacatacttc ccccagttcg ggagacccaa
     1801 gtcaactcct gagatccgtt cgtgcacatc gggaaggtga acttcaacaa aatcctcagc
     1861 ctcgtcaccg ttcttgaaaa cggtagacgg gtctgccagg gggtgcataa gggggataac
     1921 cgaggatacc aacaactcca tatgttgctg gaggagggat gattgcatct ctgggattaa
     1981 gacattgccg tcaggtccta atattatacc attgaaaaat accccgttta catgaggatg
     2041 acacctcccc ccaactctta aacacccttt tgaagggatg atctcattcc aagttctgac
     2101 tgacttgtag tgagcatcgg cttccatcaa ggtcttgttg aatatggtat atgcttttcc
     2161 aaacccaggg acaagttttc ttaaatgact gagacgtctg aaactcactg acttggtggt
     2221 catgatggac tctagtgcat ccagacactc ctctctcttc ttgaccaact cctctacaac
     2281 aaggtgctca atttcgtctg agcgaaagtc gtgcaaattc accaactgac cgggagggca
     2341 ccatttggtt tcatttgatg tttgcatcgc gacccatgtt ccatccataa gtctaagtcc
     2401 tagaactcca cataacttga gtttgcatgc tccttttaaa gacttatata ggcctctttc
     2461 atctacaaag ccgcaagtct cactcccttt ggatgctctc ttccctctac tattggtaaa
     2521 aatgtcacaa gacatcccta gtctcggatt ctcgggcatc caaatggtgt aatcgtggtt
     2581 agtggagcag taggtagaag acaccgctac tcctgagcaa ttcccgccag ggaagaccgg
     2641 cgagtgaagg gatctgtcat atgggtccaa atctgccaca cttggagata tgataacgag
     2701 agactccttg gtggttttta cagttcgaag ccagtggtag tcagggtacg gattgtgtag
     2761 agactcttca tatctggggt caccggccat cttccagttg tacgcggctc tacatgcatc
     2821 tggtgttggg cggaaatgct ttcttttgaa cgtggttgtg acataaccaa cgaagttagt
     2881 gtaggtttca gcctccgtca caacgcctgt gcaagtgaac ccgttcattt ttatggctga
     2941 gatgtatcca actttaagtt ccatgtagga gaaccctgac aggttggtgc atccttcgtc
     3001 ctccactacc aaattgtttg ggcagctgag gtgatgtatg tcaatcgggc tccagggacc
     3061 aagcttgtct ggtatcgtgt aaatagggaa tttcccaaaa cacaatggaa aaaccagaag
     3121 gggtacaaac aggagagcct gaggaaccat ggtggcggcg ctagcataac ttcgtataaa
     3181 gtatcctata cgaagttatt tgccttaacc cagaaattat cactgttatt ctttagaatg
     3241 gtgcaaagaa taacttcgta taatgtatgc tatacgaagt tatgaattcg atatcaagct
     3301 tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     3361 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     3421 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     3481 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     3541 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     3601 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     3661 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
     3721 gctgctcgcc tatgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     3781 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     3841 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg
     3901 ataccgagcg ctgctcgaga gatctacggg tggcatccct gtgacccctc cccagtgcct
     3961 ctcctggccc tggaagttgc cactccagtg cccaccagcc ttgtcctaat aaaattaagt
     4021 tgcatcattt tgtctgacta ggtgtccttc tataatatta tggggtggag gggggtggta
     4081 tggagcaagg ggcaagttgg gaagacaacc tgtagggcct gcggggtcta ttgggaacca
     4141 agctggagtg cagtggcaca atcttggctc actgcaatct ccgcctcctg ggttcaagcg
     4201 attctcctgc ctcagcctcc cgagttgttg ggattccagg catgcatgac caggctcagc
     4261 taatttttgt ttttttggta gagacggggt ttcaccatat tggccaggct ggtctccaac
     4321 tcctaatctc aggtgatcta cccaccttgg cctcccaaat tgctgggatt acaggcgtga
     4381 accactgctc ccttccctgt ccttctgatt ttgtaggtaa ccacgtgcgg accgagcggc
     4441 cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
     4501 aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg
     4561 agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt
     4621 gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt
     4681 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     4741 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     4801 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     4861 caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     4921 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     4981 aacactcaac cctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggc
     5041 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     5101 aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     5161 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     5221 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     5281 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     5341 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     5401 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     5461 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     5521 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     5581 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     5641 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     5701 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     5761 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     5821 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     5881 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     5941 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     6001 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     6061 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     6121 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     6181 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     6241 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     6301 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     6361 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     6421 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     6481 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     6541 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     6601 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     6661 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     6721 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     6781 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     6841 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     6901 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     6961 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     7021 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     7081 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     7141 tttacggttc ctggcctttt gctggccttt tgctcacatg t
//
