LOCUS       Exported                5676 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  AAV expression of EGFP from tMCK promoter.
ACCESSION   .
VERSION     .
KEYWORDS    pENN.AAV.tMCK.PI.eGFP.WPRE.bGH
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5676)
  TITLE     AAV plasmids
REFERENCE   2  (bases 1 to 5676)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5676
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     intron          1004..1136
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1119..1138
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1148..1157
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1154..1873
                     /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             1154..1870
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1199..1220)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1460..1479)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1807..1828
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1940..1960)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2358..2369)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2436..2453)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2442..2649
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2737..2866
                     /label=AAV2 ITR
     primer_bind     complement(2885..2902)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2885..2901)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2894..2916)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3043..3498
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3130..3149)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3340..3361
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3524..3628
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3629..4489
                     /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(3847..3866)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4660..5248
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5149..5168
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5402..5419
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5536..5557
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5572..5602
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5610..5626
                     /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     5615..5637
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5634..5650
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5634..5650
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ccttaattag gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcggc
       61 ctcagtgagc gagcgagcgc gcagagaggg agtggccaac tccatcacta ggggttcctt
      121 gtagttaatg attaacccgc catgctactt atctacgtag ccatgctcta ggaagatcgg
      181 aattcgccct taagctagcc actacgggtc taggctgccc atgtaaggag gcaaggcctg
      241 gggacacccg agatgcctgg ttataattaa ccccaacacc tgctgccccc ccccccccaa
      301 cacctgctgc ctgagcctga gcggttaccc caccccggtg cctgggtctt aggctctgta
      361 caccatggag gagaagctcg ctctaaaaat aaccctgtcc ctggtgggcc cactacgggt
      421 ctaggctgcc catgtaagga ggcaaggcct ggggacaccc gagatgcctg gttataatta
      481 accccaacac ctgctgcccc ccccccccca acacctgctg cctgagcctg agcggttacc
      541 ccaccccggt gcctgggtct taggctctgt acaccatgga ggagaagctc gctctaaaaa
      601 taaccctgtc cctggtgggc cactacgggt ctaggctgcc catgtaagga ggcaaggcct
      661 ggggacaccc gagatgcctg gttataatta accccaacac ctgctgcccc ccccccccca
      721 acacctgctg cctgagcctg agcggttacc ccaccccggt gcctgggtct taggctctgt
      781 acaccatgga ggagaagctc gctctaaaaa taaccctgtc cctggtgggc ccctccctgg
      841 ggacagcccc tcctggctag tcacaccctg taggctcctc tatataaccc aggggcacag
      901 gggctgcccc cgggtcacgg ggatcctcta gacctccgaa cggtaagagc ctagcatgta
      961 gaactggtta ccctgcagaa gttggtcgtg aggcactggg caggtaagta tcaaggttac
     1021 aagacaggtt taaggagacc aatagaaact gggcttgtcg agacagagaa gactcttgcg
     1081 tttctgatag gcacctattg gtcttactga catccacttt gcctttctct ccacaggtgt
     1141 ccaggcggcc gccatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     1201 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
     1261 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     1321 gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc
     1381 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     1441 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     1501 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     1561 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
     1621 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
     1681 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     1741 gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg
     1801 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     1861 gctgtacaag taataagctt ggatccaatc aacctctgga ttacaaaatt tgtgaaagat
     1921 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     1981 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     2041 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     2101 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     2161 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     2221 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     2281 aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt
     2341 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     2401 cggctctgcg gcctcttccg cgtcttcgag atctgcctcg actgtgcctt ctagttgcca
     2461 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     2521 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     2581 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     2641 tgctggggac tcgagttaag ggcgaattcc cgataaggat cttcctagag catggctacg
     2701 tagataagta gcatggcggg ttaatcatta actacaagga acccctagtg atggagttgg
     2761 ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac
     2821 gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagcctt aattaaccta
     2881 attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta
     2941 atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg
     3001 atcgcccttc ccaacagttg cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg
     3061 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     3121 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     3181 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     3241 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     3301 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     3361 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     3421 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     3481 tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     3541 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     3601 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     3661 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     3721 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     3781 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     3841 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     3901 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     3961 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     4021 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     4081 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     4141 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     4201 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     4261 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     4321 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     4381 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     4441 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     4501 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     4561 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     4621 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     4681 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     4741 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     4801 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     4861 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     4921 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     4981 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     5041 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     5101 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     5161 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     5221 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     5281 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     5341 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     5401 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     5461 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     5521 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     5581 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     5641 cagctatgac catgattacg ccagatttaa ttaagg
//
