LOCUS       Exported                7794 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    rAAV-syn::FLEX-rev::PSAMQ79G,Q139G:5HT3HC-IRES-GFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7794)
  AUTHORS   Magnus CJ, Lee PH, Atasoy D, Su HH, Looger LL, Sternson SM
  TITLE     Chemical and genetic engineering of selective ion channel-ligand 
            interactions.
  JOURNAL   Science. 2011 Sep 2;333(6047):1292-6.
  PUBMED    21885782
REFERENCE   2  (bases 1 to 7794)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7794
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   64..193
                     /label=AAV2 ITR
     polyA_signal    355..476
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(392..411)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     446..465
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    554..1142
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             660..671
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     1069..1089
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    complement(1214..1247)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     protein_bind    complement(1318..1351)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites (Lee and 
                     Saito, 1988)."
     misc_feature    2869..3420
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3013..3030)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             3444..4160
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(3486..3507)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3747..3766)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4094..4115
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     intron          4205..4337
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     protein_bind    4348..4381
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     protein_bind    4444..4477
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites (Lee and 
                     Saito, 1988)."
     intron          4490..4622
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     repeat_region   5215..5344
                     /label=AAV2 ITR
     rep_origin      5416..5871
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5503..5522)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5713..5734
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5897..6001
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6002..6862
                     /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(6220..6239)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7033..7621
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7522..7541
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7775..7792
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
       61 cagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc
      121 tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc
      181 actaggggtt ccttgtagtt aatgattaac ccgccatgct acttatctac gtagccatgc
      241 tctaggaaga tccttatcga ttttaccaca tttgtagagg ttttacttgc tttaaaaaac
      301 ctcccacatc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
      361 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
      421 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
      481 gtctgctcga agcggccgcc cgggtcgact ctagatgcat gctcgagcgg ccgcccgggt
      541 cgacggtatc gatgcgggga ggcggcccaa agggagatcc gactcgtctg agggcgaagg
      601 cgaagacgcg gaagaggccg cagagccggc agcaggccgc gggaaggaag gtccgctgga
      661 ttgagggccg aagggacgta gcagaaggac gtcccgcgca gaatccaggt ggcaacacag
      721 gcgagcagcc aaggaaagga cgatgatttc cccgacaaca ccacggaatt gtcagtgccc
      781 aacagccgag cccctgtcca gcagcgggca aggcaggcgg cgatgagttc cgccgtggca
      841 atagggaggg ggaaagcgaa agtcccggaa aggagctgac aggtggtggc aatgccccaa
      901 ccagtggggg ttgcgtcagc aaacacagtg cacaccacgc cacgttgcct gacaacgggc
      961 cacaactcct cataaagaga cagcaaccag gatttataca aggaggagaa aatgaaagcc
     1021 atacgggaag caatagcatg atacaaaggc attaaagcag cgtatccaca tagcgtaaaa
     1081 ggagcaacat agttaagaat accagtcaat ctttcacaaa ttttgtaatc cagaggttga
     1141 ttatcgataa gcttctcgag tggatcatct gcagaattcg cccttttcag tgtttcgaac
     1201 ctcttcgagg gacataactt cgtataatgt atgctatacg aagttatggg tcgatggtga
     1261 gatctggact agagggtcga tggtgatgct tggatcaaaa taggaagaat cccaatgata
     1321 acttcgtata aagtatccta tacgaagtta tactagcgcg ccagctggat agcttgaatt
     1381 caccatgcgc tgttctccag gcggcgtgtg gctcgccctg gctgcttccc ttctgcacgt
     1441 tagcctgcag ggtgagttcc agcgcaaact gtataaggag cttgttaaga attataaccc
     1501 cctggagcgg ccggtcgcaa atgattccca gccactgaca gtgtacttca gcctctcctt
     1561 gctgcagatc atggacgtgg atgaaaagaa ccaggtgctg accactaata tttggttggg
     1621 catgtcctgg accgatcact acttgcagtg gaatgtgagc gaatacccag gtgtaaagac
     1681 tgtaagattc cctgacggcc aaatctggaa accagatatc ctgctgtaca acagcgcaga
     1741 cgaaaggttt gatgcaacat ttcacaccaa cgtgttggtc aattcttcag gccactgcgg
     1801 ctacctgccc cctggaatct tcaagtcctc atgctatatc gacgtccgct ggtttccctt
     1861 cgacgtccag cactgcaaac tcaaattcgg gagctggagc tacggcggat ggagcctgga
     1921 tctgcaaatg caggaggctg acatctctgg ttacatcccg aatggggagt gggaccttgt
     1981 gggaatcccc ggtaaaagaa gcgagcgatt ttatgaatgc tgcaaggaac cctaccctga
     2041 cgtaacattc acagttatca tcagaagaag gccattgttc tacgccgtta gtttgttgct
     2101 ccccagtatt tttctcatgg tcgtggacat cgtgggattt tgtctcccac ctgatagcgg
     2161 ggagagggtc tcctttaaga ttaccttgtt gctcggctat tctgtatttc tgatcatcgt
     2221 gtccgatacc cttcctgcca caatcggcac tccgctgata ggagtgtatt tcgtcgtgtg
     2281 tatggcactc ctggtgataa gtctggcgga aactatcttc attgtacggc tggtacataa
     2341 gcaggacctg caaagacccg tgccagactg gttgcgacac cttgtgctgg acagaattgc
     2401 atggattctg tgtcttggcg agcaacctat ggcccaccgg ccacctgcaa cctttcaagc
     2461 caacaagaca gacgattgta gtgggtctga tctgttgcct gctatgggga atcactgctc
     2521 ccatgttggg ggaccacaag atttggaaaa gaccccacgg gggcggggat caccccttcc
     2581 tcctccccga gaagcctctc tcgctgtccg ggggctgctc caggaactgt caagcatccg
     2641 acattttctg gagaagcagg acgagatgga cgaagtcgct gccgactggc tgcgagtggg
     2701 ctacgtcctt gacaggctgc tgtttcggat ctacttgctg gcggtgctgg cttattccat
     2761 tactctggtg acactctggt ccatatggca ctacagttag gcggccgcct cgagtctaga
     2821 tctccggacc cgggctagcg caattgcggc cgcaattccc cccscctaac gttactggcc
     2881 gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gttattttcc accatattgc
     2941 cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg agcattccta
     3001 ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag
     3061 ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc aggcagcgga
     3121 accccccacc tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa gatacacctg
     3181 caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa agagtcaaat
     3241 ggctctccta agcgtattca acaaggggct gaaggatgcc cagaaggtac cccattgtat
     3301 gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa
     3361 acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac gatgataata
     3421 tggccacaac catgggggat ccggtgagca agggcgagga gctgttcacc ggggtggtgc
     3481 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     3541 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     3601 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
     3661 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     3721 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     3781 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     3841 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     3901 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
     3961 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     4021 tgctgctgcc cgacaaccac tacctgagca cccagtccaa gctgagcaaa gaccccaacg
     4081 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     4141 tggacgagct gtacaagtaa tctagagtcg aggtaccgat gaatagctaa ggtcgaggcc
     4201 gcaggtaagt atcaaggtta caagacaggt ttaaggagac caatagaaac tgggcttgtc
     4261 gagacagaga agactcttgc gtttctgata ggcacctatt ggtcttactg acatccactt
     4321 tgcctttctc tccacaggtg tcgaagtata acttcgtata gcatacatta tacgaagtta
     4381 tgtgtcgatg gtgatgcttg gcaattcggg tcgatggtga agcattggtc ttcctatttt
     4441 gatataactt cgtataggat actttatacg aagttatggg gatcatcacc tgtggagaga
     4501 aaggcaaagt ggatgtcagt aagaccaata ggtgcctatc agaaacgcaa gagtcttctc
     4561 tgtctcgaca agcccagttt ctattggtct ccttaaacct gtcttgtaac cttgatactt
     4621 acctgatcca acggctgcga cttggggcag ggggtcctag gggtggtctg gccaggagcc
     4681 gcgggggcgg actgcgcggt gcccaggagc acagctgcgc tctcaggcac gacacgactc
     4741 ctccgctgcc caccgcagac tgaggcagcg ctgagtcgcc ggcgccgcag cgcagatggt
     4801 cgcgcccgtg cccccctatc tcgcgcctcg cgtggtgcgg tccggctggg ccggcggcgg
     4861 cgcggacgcg accaaggtgg ccgggaaggg gagtttgcgg gggaccggcg agtgacgtca
     4921 gcgcgccttc agtgctgagg cggcggtggc gcgcgccgcc aggcgggggc gaaggcactg
     4981 tccgcggtgc tgaagctggc agtgcgcacg cgcctcgccg catcctgttt cccctccccc
     5041 tctctgatag gggatgcgca atttggggaa tgggggttgg gtgcttgtcc agtgggtcgg
     5101 ggtcggtcgt caatttaaat actgaaaagg gcgaattcca gcacactgga gctcggatct
     5161 tcctagagca tggctacgta gataagtagc atggcgggtt aatcattaac tacaaggaac
     5221 ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc
     5281 gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc
     5341 gcagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     5401 tgaatggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     5461 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     5521 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     5581 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     5641 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     5701 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     5761 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     5821 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac
     5881 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     5941 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     6001 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     6061 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     6121 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     6181 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     6241 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     6301 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     6361 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     6421 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     6481 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     6541 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     6601 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     6661 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     6721 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     6781 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     6841 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     6901 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     6961 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     7021 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     7081 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     7141 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     7201 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     7261 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     7321 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     7381 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     7441 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     7501 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     7561 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     7621 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     7681 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     7741 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg agga
//
