LOCUS       Exported                6246 bp DNA     circular SYN 16-JUN-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 6246)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Jun 20, 2022 from SnapGene Viewer 6.0.5
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6246
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     rep_origin      1..141
                     /direction=RIGHT
                     /label=ITR
     promoter        230..921
                     /label=GFAP(0.7Kb)
     CDS             971..1012
                     /codon_start=1
                     /label=Lyn_tail
                     /translation="MGCIKSKRKDGRIP"
     CDS             1013..1720
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1158..1176)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1426..1445
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1733..2449
                     /codon_start=1
                     /product="pH-sensitive mutant of green fluorescent protein 
                     (Ng et al., 2002)"
                     /label=superecliptic pHluorin
                     /note="excited strongly by blue light at neutral pH but 
                     minimally fluorescent at acidic pH"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNDHQVYIMADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLFTTSTLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(1763..1791)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2386..2406
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     rep_origin      2540..3131
                     /direction=RIGHT
                     /label=WPRE
     rep_origin      3197..3424
                     /direction=RIGHT
                     /label=bGHpA
     rep_origin      3509..3649
                     /direction=RIGHT
                     /label=ITR
     rep_origin      4566..5423
                     /direction=RIGHT
                     /label=Amp(R)
     rep_origin      5574..6241
                     /direction=RIGHT
                     /label=pUCori
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggcaatt cagtcgataa ctataacggt cctaaggtag
      181 cgatttaaat acgcgctctc ttaaggtagc cccgggacgc gtcaattgag aacatatcct
      241 ggtgtggagt aggggacgct gctctgacag aggctcgggg gcctgagctg gctctgtgag
      301 ctggggagga ggcagacagc caggccttgt ctgcaagcag acctggcagc attgggctgg
      361 ccgcccccca gggcctcctc ttcatgccca gtgaatgact caccttggca cagacacaat
      421 gttcggggtg ggcacagtgc ctgcttcccg ccgcacccca gcccccctca aatgccttcc
      481 gagaagccca ttgagcaggg ggcttgcatt gcaccccagc ctgacagcct ggcatcttgg
      541 gataaaagca gcacagcccc ctaggggctg cccttgctgt gtggcgccac cggcggtgga
      601 gaacaaggct ctattcagcc tgtgcccagg aaaggggatc aggggatgcc caggcatgga
      661 cagtgggtgg caggggggga gaggagggct gtctgcttcc cagaagtcca aggacacaaa
      721 tgggtgaggg gactggaggg gagctctccc catagctggg ctgcggccca accccacccc
      781 ctcaggctat gccagggggt gttgccaggg gcacccgggc atcgccagtc tagcccactc
      841 cttcataaag ccctcgcatc ccaggagcga gcagagccag agcaggttgg agaggagacg
      901 catcacctcc gctgctcgcc gctagcgttt aaacttaagc ttggtaccga gctcggatct
      961 cgagagaaat atgggatgta ttaaatcaaa aaggaaagac gggagaattc ccatggtgag
     1021 caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca aggtgcacat
     1081 ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta
     1141 cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc ccttcgcctg
     1201 ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc accccgccga
     1261 catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa
     1321 cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg acggcgagtt
     1381 catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg taatgcagaa
     1441 gaagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg gcgccctgaa
     1501 gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa
     1561 gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg tcaacatcaa
     1621 gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg aacgcgccga
     1681 gggccgccac tccaccggcg gcatggacga gctgtacaag ggatccgcca ccatgagtaa
     1741 aggagaagaa cttttcactg gagttgtccc aattcttgtt gaattagatg gtgatgttaa
     1801 tgggcacaaa ttttctgtca gtggagaggg tgaaggtgat gcaacatacg gaaaacttac
     1861 ccttaaattt atttgcacta ctggaaaact acctgttcct tggccaacac ttgtcactac
     1921 tttaacttat ggtgttcaat gcttttcaag atacccagat catatgaaac ggcatgactt
     1981 tttcaagagt gccatgcccg aaggttatgt acaggaaaga actatatttt tcaaagatga
     2041 cgggaactac aagacacgtg ctgaagtcaa gtttgaaggt gatacccttg ttaatagaat
     2101 cgagttaaaa ggtattgatt ttaaagaaga tggaaacatt cttggacaca aattggaata
     2161 caactataac gatcaccagg tgtacatcat ggcagacaaa caaaagaatg gaatcaaagc
     2221 taacttcaaa attagacaca acattgaaga tggaggcgtt caactagcag accattatca
     2281 acaaaatact ccaattggcg atgggcccgt ccttttacca gacaaccatt acctgtttac
     2341 aacttctact ctttcgaaag atcccaacga aaagagagac cacatggtcc ttcttgagtt
     2401 tgtaacagct gctgggatta cacatggcat ggatgaacta tacaaataaa gcggccgctc
     2461 gagtctagag ggcccttcga aggtaagcct atccctaacc ctctcctcgg tctcgattct
     2521 acgcgtaccg gttaatcgat aatcaacctc tggattacaa aatttgtgaa agattgactg
     2581 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     2641 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     2701 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     2761 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     2821 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     2881 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat
     2941 cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct
     3001 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
     3061 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
     3121 cctccccgca tcgaaacccg ctgatcagcc ggtcatcatc accatcacca ttgagtttaa
     3181 acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     3241 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     3301 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     3361 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
     3421 atggcttctg aggcggaaag aaccagatcc tctcttaagg tagcatcgag atttaaatta
     3481 gggataacag ggtaatggcg cgggccgcag gaacccctag tgatggagtt ggccactccc
     3541 tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc
     3601 tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg
     3661 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat
     3721 agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     3781 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     3841 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     3901 ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg
     3961 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     4021 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcgggctat tcttttgatt
     4081 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     4141 ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa
     4201 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     4261 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     4321 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     4381 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     4441 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     4501 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     4561 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     4621 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     4681 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     4741 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     4801 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     4861 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     4921 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     4981 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     5041 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     5101 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     5161 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     5221 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     5281 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     5341 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     5401 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     5461 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     5521 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     5581 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     5641 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     5701 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     5761 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     5821 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     5881 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     5941 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     6001 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     6061 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     6121 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     6181 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     6241 acatgt
//
