LOCUS       Exported                6072 bp ds-DNA     circular SYN 25-JUL-2019
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 6072)
  TITLE     AAV plasmids
REFERENCE   2  (bases 1 to 6072)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Apr 21, 2020 from SnapGene 4.3.4
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6072
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(23..34)
                     /label=L4440
                     /note="L4440 vector, forward primer"
                     /note="color: black; sequence: AGCGAGTCAGTGAGCGAG"
     primer_bind     62..73
                     /label=L4440
                     /note="L4440 vector, forward primer"
                     /note="color: black; sequence: AGCGAGTCAGTGAGCGAG"
     intron          1004..1136
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
                     /note="color: #------"
     primer_bind     1119..1138
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
                     /note="color: black; sequence: TTGCCTTTCTCTCCACAGGT"
     primer_bind     1126..1171
                     /label=F198
                     /note="color: black; sequence: 
                     tctctccacaggtgtccaggcggccgcgccaccatggagacagacacactc; added:
                     2019-07-17"
     primer_bind     1139..1171
                     /label=F183
                     /note="color: black; sequence: 
                     gtccaggcggccgcgccaccatggagacagacacactc; added: 2019-07-03"
     regulatory      1148..1153
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
     regulatory      1154..1156
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
     sig_peptide     1157..1216
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /note="color: #993366
                     Cleavage site after base 1213"
     CDS             1217..1243
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="color: #cc99b2"
                     /translation="YPYDVPDYA"
     misc_feature    1250..2212
                     /label=TurboID
                     /note="color: #ff0000; direction: RIGHT"
     primer_bind     complement(1871..1883)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
                     /note="color: black; sequence: ATAATACCGCGCCACATAGC"
     CDS             2213..2254
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /note="color: #cc99b2"
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(2242..2305)
                     /label=R198
                     /note="color: black; sequence: 
                     attttgtaatccagaggttgattggatccaagcttttagagttcatctttggtgctgtc
                     cagg; added: 2019-07-17"
     primer_bind     complement(2242..2282)
                     /label=R183
                     /note="color: black; sequence: 
                     ggatccaagcttttagagttcatctttggtgctgtccagg; added: 
                     2019-07-03"
     misc_feature    2255..2266
                     /label=KDEL
                     /note="color: #993366"
     primer_bind     complement(2336..2356)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
                     /note="color: black; sequence: CATAGCGTAAAAGGAGCAACA"
     CDS             complement(2754..2765)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /note="color: #cc99b2
                     Cleavage site after base 2753"
                     /translation="IEGR"
     primer_bind     complement(2832..2849)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
                     /note="color: black; sequence: TAGAAGGCACAGTCGAGG"
     polyA_signal    2838..3045
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
                     /note="color: #a6acb3"
     repeat_region   3133..3262
                     /label=AAV2 ITR
                     /note="color: #ffcc99"
     primer_bind     complement(3179..3190)
                     /label=L4440
                     /note="L4440 vector, forward primer"
                     /note="color: black; sequence: AGCGAGTCAGTGAGCGAG"
     primer_bind     3240..3251
                     /label=L4440
                     /note="L4440 vector, forward primer"
                     /note="color: black; sequence: AGCGAGTCAGTGAGCGAG"
     primer_bind     complement(3281..3298)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
                     /note="color: black; sequence: TGTAAAACGACGGCCAGT"
     primer_bind     complement(3281..3297)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     primer_bind     complement(3290..3312)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
                     /note="color: black; sequence: CCCAGTCACGACGTTGTAAAACG"
     rep_origin      3439..3894
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     primer_bind     complement(3526..3545)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
                     /note="color: black; sequence: AGGGAAGAAAGCGAAAGGAG"
     primer_bind     3736..3757
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
                     /note="color: black; sequence: GTGGACTCTTGTTCCAAACTGG"
     promoter        3920..4024
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     CDS             4025..4885
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 4025 .. 4093 / #ccffcc / signal sequence
                        2: 4094 .. 4885 / #ccffcc
                     Cleavage site after base 4093"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4243..4262)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
                     /note="color: black; sequence: ATAATACCGCGCCACATAGC"
     rep_origin      5056..5644
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     primer_bind     5545..5564
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
                     /note="color: black; sequence: GGGAAACGCCTGGTATCTTT"
     primer_bind     5798..5815
                     /label=L4440
                     /note="L4440 vector, forward primer"
                     /note="color: black; sequence: AGCGAGTCAGTGAGCGAG"
     protein_bind    5932..5953
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
                     /note="color: #31849b"
     promoter        5968..5998
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This forward directional feature has 3 segments:
                        1: 5968 .. 5973 / #ffffff / -35
                        2: 5974 .. 5991 / #ffffff
                        3: 5992 .. 5998 / #ffffff / -10"
     protein_bind    6006..6022
                     /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)."
                     /note="color: #31849b"
     primer_bind     6011..6033
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
                     /note="color: black; sequence: AGCGGATAACAATTTCACACAGG"
     primer_bind     6030..6046
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: RIGHT"
     primer_bind     6030..6046
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
                     /note="color: black; sequence: CAGGAAACAGCTATGAC"
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 gccatggaga cagacacact cctgctatgg gtgctgctgc tctgggttcc
     1201 aggttccact ggtgactacc catacgatgt tccagattac gctgctagca aagacaatac
     1261 tgtgcctctg aagctgatcg ctctcctggc taatggcgag ttccatagtg gcgaacagct
     1321 gggagaaacc ctgggcatgt ccagggccgc tatcaacaag cacattcaga ctctgcgcga
     1381 ctggggcgtg gacgtgttca ccgtgcccgg aaagggctac tctctgcccg agcctatccc
     1441 gctgctgaac gctaaacaga ttctgggaca gctggacggc gggagcgtgg cagtcctgcc
     1501 tgtggtcgac tccaccaatc agtacctgct ggatcgaatc ggcgagctga agagtgggga
     1561 tgcttgcatt gcagaatatc agcaggcagg gagaggaagc agagggagga aatggttctc
     1621 tccttttgga gctaacctgt acctgagtat gttttggcgc ctgaagcggg gaccagcagc
     1681 aatcggcctg ggcccggtca tcggaattgt catggcagaa gcgctgcgaa agctgggagc
     1741 agacaaggtg cgagtcaaat ggcccaatga cctgtatctg caggatagaa agctggcagg
     1801 catcctggtg gagctggccg gaataacagg cgatgctgca cagatcgtca ttggcgccgg
     1861 gattaacgtg gctatgaggc gcgtggagga aagcgtggtc aatcagggct ggatcacact
     1921 gcaggaagca gggattaacc tggacaggaa tactctggcc gctacgctga tccgagagct
     1981 gcgggcagcc ctggaactgt tcgagcagga aggcctggct ccatatctgc cacggtggga
     2041 gaagctggat aacttcatca atagacccgt gaagctgatc attggggaca aagagatttt
     2101 cgggattagc cgggggattg ataaacaggg agccctgctg ctggaacagg acggagttat
     2161 caaaccctgg atgggcggag aaatcagtct gcggtctgcc gaaaagctgc agggcaagcc
     2221 catccccaac cccctgctgg gcctggacag caccaaagat gaactctaat aagcttggat
     2281 ccaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     2341 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     2401 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
     2461 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
     2521 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
     2581 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     2641 tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc
     2701 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
     2761 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
     2821 ttcgagatct gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     2881 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     2941 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     3001 cagcaagggg gaggattggg aagacaatag caggcatgct ggggactcga gttaagggcg
     3061 aattcccgat aaggatcttc ctagagcatg gctacgtaga taagtagcat ggcgggttaa
     3121 tcattaacta caaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     3181 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
     3241 cagtgagcga gcgagcgcgc agccttaatt aacctaattc actggccgtc gttttacaac
     3301 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     3361 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     3421 gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     3481 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     3541 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     3601 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     3661 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     3721 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     3781 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     3841 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg
     3901 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     3961 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     4021 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     4081 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     4141 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     4201 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     4261 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     4321 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     4381 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     4441 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     4501 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     4561 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     4621 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     4681 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     4741 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     4801 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     4861 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     4921 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     4981 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     5041 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     5101 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     5161 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     5221 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     5281 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     5341 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     5401 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     5461 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     5521 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     5581 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     5641 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     5701 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     5761 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     5821 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     5881 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
     5941 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
     6001 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccag
     6061 atttaattaa gg
//
