LOCUS       Exported                6003 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 6003)
  TITLE     AAV plasmids
REFERENCE   2  (bases 1 to 6003)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Apr 21, 2020 from SnapGene 4.3.4
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6003
                     /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"
     regulatory      1148..1153
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
     regulatory      1154..1157
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
     CDS             1157..1198
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /note="color: #cc99b2"
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(1826..1838)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
                     /note="color: black; sequence: ATAATACCGCGCCACATAGC"
     CDS             2168..2194
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /note="color: #993366"
                     /translation="LPPLERLTL"
     primer_bind     complement(2180..2240)
                     /label=R199
                     /note="color: black; sequence: 
                     acaaattttgtaatccagaggttgattggatccaagcttttagtccagggtcaggcgct
                     c; added: 2019-07-17"
     primer_bind     complement(2267..2287)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
                     /note="color: black; sequence: CATAGCGTAAAAGGAGCAACA"
     CDS             complement(2685..2696)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /note="color: #cc99b2
                     Cleavage site after base 2684"
                     /translation="IEGR"
     primer_bind     complement(2763..2780)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
                     /note="color: black; sequence: TAGAAGGCACAGTCGAGG"
     polyA_signal    2769..2976
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
                     /note="color: #a6acb3"
     repeat_region   3064..3193
                     /label=AAV2 ITR
                     /note="color: #ffcc99"
     primer_bind     complement(3110..3121)
                     /label=L4440
                     /note="L4440 vector, forward primer"
                     /note="color: black; sequence: AGCGAGTCAGTGAGCGAG"
     primer_bind     3171..3182
                     /label=L4440
                     /note="L4440 vector, forward primer"
                     /note="color: black; sequence: AGCGAGTCAGTGAGCGAG"
     primer_bind     complement(3212..3229)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
                     /note="color: black; sequence: TGTAAAACGACGGCCAGT"
     primer_bind     complement(3212..3228)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     primer_bind     complement(3221..3243)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
                     /note="color: black; sequence: CCCAGTCACGACGTTGTAAAACG"
     rep_origin      3370..3825
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     primer_bind     complement(3457..3476)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
                     /note="color: black; sequence: AGGGAAGAAAGCGAAAGGAG"
     primer_bind     3667..3688
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
                     /note="color: black; sequence: GTGGACTCTTGTTCCAAACTGG"
     promoter        3851..3955
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     CDS             3956..4816
                     /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: 3956 .. 4024 / #ccffcc / signal sequence
                        2: 4025 .. 4816 / #ccffcc
                     Cleavage site after base 4024"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4174..4193)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
                     /note="color: black; sequence: ATAATACCGCGCCACATAGC"
     rep_origin      4987..5575
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     primer_bind     5476..5495
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
                     /note="color: black; sequence: GGGAAACGCCTGGTATCTTT"
     primer_bind     5729..5746
                     /label=L4440
                     /note="L4440 vector, forward primer"
                     /note="color: black; sequence: AGCGAGTCAGTGAGCGAG"
     protein_bind    5863..5884
                     /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        5899..5929
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This forward directional feature has 3 segments:
                        1: 5899 .. 5904 / #ffffff / -35
                        2: 5905 .. 5922 / #ffffff
                        3: 5923 .. 5929 / #ffffff / -10"
     protein_bind    5937..5953
                     /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     5942..5964
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
                     /note="color: black; sequence: AGCGGATAACAATTTCACACAGG"
     primer_bind     5961..5977
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: RIGHT"
     primer_bind     5961..5977
                     /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 gccatgggca agcccatccc caaccccctg ctgggcctgg acagcaccgc
     1201 tagcaaagac aatactgtgc ctctgaagct gatcgctctc ctggctaatg gcgagttcca
     1261 tagtggcgaa cagctgggag aaaccctggg catgtccagg gccgctatca acaagcacat
     1321 tcagactctg cgcgactggg gcgtggacgt gttcaccgtg cccggaaagg gctactctct
     1381 gcccgagcct atcccgctgc tgaacgctaa acagattctg ggacagctgg acggcgggag
     1441 cgtggcagtc ctgcctgtgg tcgactccac caatcagtac ctgctggatc gaatcggcga
     1501 gctgaagagt ggggatgctt gcattgcaga atatcagcag gcagggagag gaagcagagg
     1561 gaggaaatgg ttctctcctt ttggagctaa cctgtacctg agtatgtttt ggcgcctgaa
     1621 gcggggacca gcagcaatcg gcctgggccc ggtcatcgga attgtcatgg cagaagcgct
     1681 gcgaaagctg ggagcagaca aggtgcgagt caaatggccc aatgacctgt atctgcagga
     1741 tagaaagctg gcaggcatcc tggtggagct ggccggaata acaggcgatg ctgcacagat
     1801 cgtcattggc gccgggatta acgtggctat gaggcgcgtg gaggaaagcg tggtcaatca
     1861 gggctggatc acactgcagg aagcagggat taacctggac aggaatactc tggccgctac
     1921 gctgatccga gagctgcggg cagccctgga actgttcgag caggaaggcc tggctccata
     1981 tctgccacgg tgggagaagc tggataactt catcaataga cccgtgaagc tgatcattgg
     2041 ggacaaagag attttcggga ttagccgggg gattgataaa cagggagccc tgctgctgga
     2101 acaggacgga gttatcaaac cctggatggg cggagaaatc agtctgcggt ctgccgaaaa
     2161 gctgcagctg cctcccctgg agcgcctgac cctggactaa taagcttgga tccaatcaac
     2221 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     2281 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     2341 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     2401 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     2461 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     2521 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     2581 ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg
     2641 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
     2701 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgagatc
     2761 tgcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     2821 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     2881 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     2941 ggaggattgg gaagacaata gcaggcatgc tggggactcg agttaagggc gaattcccga
     3001 taaggatctt cctagagcat ggctacgtag ataagtagca tggcgggtta atcattaact
     3061 acaaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
     3121 aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg
     3181 agcgagcgcg cagccttaat taacctaatt cactggccgt cgttttacaa cgtcgtgact
     3241 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     3301 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     3361 gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     3421 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     3481 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     3541 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     3601 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     3661 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     3721 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     3781 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg
     3841 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     3901 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     3961 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     4021 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     4081 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     4141 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     4201 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     4261 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     4321 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     4381 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     4441 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     4501 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     4561 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     4621 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     4681 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     4741 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     4801 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     4861 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     4921 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     4981 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     5041 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     5101 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     5161 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     5221 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     5281 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     5341 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     5401 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     5461 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     5521 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     5581 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     5641 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     5701 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     5761 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     5821 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
     5881 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
     5941 gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca gatttaatta
     6001 agg
//
