LOCUS       Exported                5504 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  AAV expression of EGFP from CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV.CMV.PI.EGFP.WPRE.bGH
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5504)
  TITLE     AAV plasmids
REFERENCE   2  (bases 1 to 5504)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5504
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..130
                     /label=AAV2 ITR
     enhancer        271..574
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        575..778
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     729..749
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          831..963
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     946..965
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      975..984
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             981..1700
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             981..1697
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1026..1047)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1287..1306)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1634..1655
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1767..1787)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2185..2196)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2263..2280)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2269..2476
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2564..2693
                     /label=AAV2 ITR
     primer_bind     complement(2712..2729)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2712..2728)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2721..2743)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2870..3325
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2957..2976)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3167..3188
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3352..3456
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3457..4317
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVVMVTTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3675..3694)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4488..5076
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4977..4996
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5230..5247
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5364..5385
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5400..5430
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5438..5454
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     5443..5465
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5462..5478
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5462..5478
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct
      181 aggaagatcg gaattcgccc ttaagctagc tagttattaa tagtaatcaa ttacggggtc
      241 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
      301 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
      361 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
      421 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
      481 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
      541 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
      601 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
      661 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
      721 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctgg
      781 tttagtgaac cgtcagatcc tgcagaagtt ggtcgtgagg cactgggcag gtaagtatca
      841 aggttacaag acaggtttaa ggagaccaat agaaactggg cttgtcgaga cagagaagac
      901 tcttgcgttt ctgataggca cctattggtc ttactgacat ccactttgcc tttctctcca
      961 caggtgtcca ggcggccgcc atggtgagca agggcgagga gctgttcacc ggggtggtgc
     1021 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     1081 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     1141 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
     1201 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     1261 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     1321 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     1381 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     1441 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
     1501 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     1561 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
     1621 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     1681 tggacgagct gtacaagtaa taagcttgga tccaatcaac ctctggatta caaaatttgt
     1741 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     1801 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     1861 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     1921 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     1981 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     2041 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     2101 tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc
     2161 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     2221 ctgctgccgg ctctgcggcc tcttccgcgt cttcgagatc tgcctcgact gtgccttcta
     2281 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     2341 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     2401 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     2461 gcaggcatgc tggggactcg agttaagggc gaattcccga ttaggatctt cctagagcat
     2521 ggctacgtag ataagtagca tggcgggtta atcattaact acaaggaacc cctagtgatg
     2581 gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc
     2641 gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagccttaat
     2701 taacctaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     2761 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     2821 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt
     2881 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     2941 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     3001 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     3061 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccccga
     3121 tagacggttt ttcgcccttt gacgctggag ttcacgttcc tcaatagtgg actcttgttc
     3181 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttt
     3241 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     3301 aacaaaatat taacgtttat aatttcaggt ggcatctttc ggggaaatgt gcgcggaacc
     3361 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     3421 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     3481 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     3541 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     3601 ctcaatagtg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     3661 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     3721 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     3781 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     3841 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     3901 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     3961 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagtaatggt aacaacgttg
     4021 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     4081 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     4141 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     4201 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     4261 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     4321 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     4381 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     4441 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     4501 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     4561 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     4621 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     4681 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     4741 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     4801 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     4861 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     4921 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     4981 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     5041 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     5101 cggttcctgg ccttttgctg cggttttgct cacatgttct ttcctgcgtt atcccctgat
     5161 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     5221 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     5281 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     5341 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     5401 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     5461 acaggaaaca gctatgacca tgattacgcc agatttaatt aagg
//
