LOCUS       Exported                6466 bp ds-DNA     circular SYN 12-JUL-2023
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6466)
  AUTHORS   William Stauffer
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Aug 13, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6466
                     /organism="synthetic DNA construct"
                     /note="color: #ffffff"
     misc_feature    1..130
                     /label=5' ITR
                     /label=nonstandard type:  ITR
                     /note="/application_notes=Allows rescue of virus from 
                     recombinant
                     plasmid and replication of the viral genome; this ITR is 
                     identical to that of the wild-type AAV2 genome."
                     /note="/description=AAV 5' inverted terminal repeat 
                     (functional
                     equivalent of wild-type 5' ITR)"
                     /note="/official_designation=5' ITR"
                     /note="/uuid=6d343a478711450d8e5c28ceb27e5b84"
                     /note="color: #6d3ed8; direction: RIGHT"
     promoter        158..1553
                     /label={L3PN.Enh001.Hsp68}
                     /label={L3PN.Enh003.Hsp68}
                     /note="/application_notes="
                     /note="/official_designation={L3PN.Enh003.Hsp68}"
                     /note="/uuid=5c7c5a2e7c89453487ba7ceeedf3163f"
                     /note="This forward directional feature has 3 segments:
                        1:  158 ..  658 / #993300 / L3PN.Enh001
                        2:  659 ..  686 / #0375bb
                        3:  687 .. 1553 / #00ccff / Hsp68 minimal promoter"
     misc_feature    1578..1583
                     /label=Kozak
                     /label=nonstandard type:  Miscellaneous
                     /note="/application_notes=Facilitates translation 
                     initiation of
                     ATG start codon downstream of the Kozak sequence."
                     /note="/description=Kozak translation initiation sequence"
                     /note="/full_name=Kozak"
                     /note="/official_designation=Kozak"
                     /note="/uuid=5f18b25a1234443494a618363f09f052"
                     /note="color: #a949ca; direction: RIGHT"
     CDS             1584..2303
                     /label=EGFP
                     /note="/application_notes=Commonly used green fluorescent
                     protein; ranked high in brightness, photostability and pH 
                     stability among all fluorescent proteins."
                     /note="/description=Enhanced green fluorescent protein; 
                     codon
                     optimized based on a variant of wild type GFP from the 
                     jellyfish Aequorea victoria"
                     /note="/full_name=EGFP reporter gene"
                     /note="/official_designation=EGFP"
                     /note="/uuid=0470849bb38e4f4b8a1fe5d436422af3"
                     /note="color: #028a02; direction: RIGHT"
     misc_signal     2335..2844
                     /label=BC.A5
                     /note="color: #000000"
     misc_feature    2875..3472
                     /label=WPRE
                     /label=nonstandard type:  Miscellaneous
                     /note="/application_notes=Enhances virus stability in 
                     packaging
                     cells, leading to higher titer of packaged virus; enhances 
                     higher expression of transgenes."
                     /note="/description=Woodchuck hepatitis virus 
                     posttranscriptional
                     regulatory element"
                     /note="/full_name=Woodchuck hepatitis virus 
                     posttranscriptional
                     regulatory element"
                     /note="/official_designation=WPRE"
                     /note="/uuid=0aedff8308474b369c622433538d7836"
                     /note="color: #946a00; direction: RIGHT"
     polyA_signal    3503..3710
                     /label=BGH pA
                     /note="/application_notes=Allows transcription termination 
                     and
                     polyadenylation of mRNA transcribed by Pol II RNA 
                     polymerase."
                     /note="/description=Bovine growth hormone polyadenylation 
                     signal"
                     /note="/full_name=Bovine growth hormone polyadenylation"
                     /note="/official_designation=BGH pA"
                     /note="/uuid=4a09734ca6c745b3aadc757c1638b651"
                     /note="color: #512bbd; direction: RIGHT"
     misc_feature    complement(3718..3847)
                     /label=3' ITR
                     /label=nonstandard type:  ITR
                     /note="/application_notes=Allows rescue of virus from 
                     recombinant
                     plasmid and replication of the viral genome; this ITR is 
                     identical to that of the wild-type AAV2 genome."
                     /note="/description=AAV 3' inverted terminal repeat"
                     /note="/official_designation=3' ITR"
                     /note="/uuid=a5d684b85c014f76a7b6d8e97945f0fb"
                     /note="color: #d84e4e; direction: LEFT"
     CDS             4775..5635
                     /label=AmpiciIIin
                     /note="/application_notes=Allows E. coli to be resistant to
                     ampiciIIin."
                     /note="/description=AmpiciIIin resistance gene"
                     /note="/full_name=Ampicillin resistance gene"
                     /note="/official_designation=Ampicillin"
                     /note="/uuid=5bcc09480c0e4c07b95ddccd8242d769"
                     /note="color: #1f36a9; direction: RIGHT"
     rep_origin      5806..6394
                     /direction=RIGHT
                     /label=pUC ori
                     /note="/application_notes=Facilitates plasmid replication 
                     in E.
                     coli; regulates high-copy plasmid number (500-700)."
                     /note="/description=pUC origin of replication"
                     /note="/full_name=pUC origin of replication"
                     /note="/official_designation=pUC ori"
                     /note="/uuid=b978ad4ff785460db448caf63b728237"
                     /note="color: #db6901"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tctagacaac tttgtataga aaagttggtc aggaaactcg ggctgaaatc
      181 ccactactgg gggaaatctg ctaacccctc tgaacctcat ttttcacttg catcaaaggg
      241 gataaagtgc ttcttgcctc tgggccgtgg gcatctgtgg gattggactg ggtcatgggg
      301 agccgtgggg tggcactggg gagctgcatc agccgccagc atctctgagg cctttgcttt
      361 ctctgaggga aggcccagtt tgatttaggt cctgagaatc ccagaatggc catttctgga
      421 aaaaataacc caaagcccag atttcccttt tagttttatc ccacacgggc cagcaacatg
      481 cataacaaaa tattccattt ctcaaaggaa gtcttagaat attggatgaa tgcactgtgt
      541 gccttgactt cattcgcaaa gcacccatgt cccttggatc ccttccgctc tccctcccgc
      601 ccccacgctg cccttgttct gggaaatcca gctctccagg agcaatcagc acagaggctc
      661 ctagtttcca tgctctagcc tgcaggcagg aacatccaaa ctgagcagcc ggggtccccc
      721 ccacccccca ccccgcccca cgcggcaact ttgagcctgt gctgggacag agcctctagt
      781 tcctaaatta gtccatgagg tcagaggcag cactgccatt gtaacgcgat tggagaggat
      841 cacgtcaccg gacacgcccc caggcatctc cctgggtctc ctaaacttgg cggggagaag
      901 ttttagccct taagttttag cctttaaccc ccatattcag aactgtgcga gttggcgaaa
      961 ccccacaaat cacaacaaac tgtacacaac accgagctag aggtgatctt tcttgtccat
     1021 tccacacagg ccttagtaat gcgtcgccat agcaacagtg tcactagtag caccagcact
     1081 tccccacacc ctccccctca ggaatccgta ctctccagtg aaccccagaa acctctggag
     1141 agttctggac aagggcggaa cccacaactc cgattactca agggaggcgg ggaagctcca
     1201 ccagacgcga aactgctgga agattcctgg ccccaaggcc tcctccggct cgctgattgg
     1261 cccagcggag agtgggcggg gccggtgaag actccttaaa ggcgcagggc ggcgagcagg
     1321 tcaccagacg ctgacagcta ctcagaacca aatctggttc catccagaga caagcgaaga
     1381 caagagaagc agagcgagcg gcgcgttccc gatcctcggc caggaccagc cttccccaga
     1441 gcatccctgc cgcggagcgc aaccttccca ggagcatccc tgccgcggag cgcaactttc
     1501 cccggagcat ccacgccgcg gagcgcagcc ttccagaagc agagcgcggc gcccaagttt
     1561 gtacaaaaaa gcaggctgcc accatggtga gcaagggcga ggagctgttc accggggtgg
     1621 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
     1681 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
     1741 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca
     1801 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     1861 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     1921 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     1981 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
     2041 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg
     2101 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     2161 ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca
     2221 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     2281 gcatggacga gctgtacaag taagtttaaa cacagggtta atcattaact acaggagcca
     2341 aggaggtcag aatactacag gtcgccgata gcattagtca acctgaccac atccctccat
     2401 cggtaaggtc cacatctagg acacagaaca gagtacatcc gcgttagtgt tattgccata
     2461 caatactagt acttgtgcga gcggaacgct acaggaatat gcgtcaagct caggtagaac
     2521 cacaagccta tcgaacgcgg tatctctaca ggtcgaatac acgccaagta cattgagttg
     2581 accattgtta attgagaact gtcccacaat catcttacct gtcttactgt ctgggaatac
     2641 tcagaatcct taggagcatg gacctaacca agtagaacgc gtaccaataa ggaaagcgca
     2701 tacggtgtga agtcacgcct aatccagtcc acacgttcgt tactgcaagc gttagataag
     2761 ctccgtcgac ctacgcctga atggatccta cctattctgc atgcgttgtg gacttcttgc
     2821 cattcgtatg tcggaatagt gagaacccag ctttcttgta caaagtggga attccgataa
     2881 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     2941 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     3001 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     3061 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     3121 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     3181 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     3241 gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc
     3301 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     3361 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     3421 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc gggaattcct
     3481 agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     3541 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     3601 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     3661 caggacagca agggggagga ttgggaagag aatagcaggc atgctgggga gggccgcagg
     3721 aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg
     3781 ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag
     3841 cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     3901 tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc
     3961 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgcct tagcgcccgc
     4021 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     4081 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     4141 acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     4201 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     4261 caactctatc tcgggctatt cttttgattt ataagggatt ttgccgattt cggtctattg
     4321 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt
     4381 tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     4441 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     4501 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     4561 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     4621 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     4681 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     4741 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     4801 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     4861 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     4921 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     4981 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     5041 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     5101 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     5161 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     5221 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     5281 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     5341 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     5401 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     5461 tgctgataaa tctggagccg gtgagcgtgg aagccgcggt atcattgcag cactggggcc
     5521 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     5581 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     5641 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     5701 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     5761 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     5821 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     5881 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     5941 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     6001 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     6061 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     6121 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     6181 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     6241 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     6301 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     6361 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     6421 gttcctggcc ttttgctggc cttttgctca catgtcctgc aggcag
//
