LOCUS       Exported                6828 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Green glucose sensor with mRuby2 fusion, cytosolic.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV.hSynap.(cyto)iGlucoSnFR-mRuby2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6828)
  AUTHORS   Looger, L et al
  TITLE     Glucose biosensors
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 6828)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6828
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   4..133
                     /label=AAV2 ITR
     promoter        209..656
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     primer_bind     1887..1908
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2014..2035)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2275..2294)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             2632..2661
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2692..3402
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTH
                     MALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAV
                     AKFAGLGGGMDELYK"
     misc_feature    3418..4006
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3471..3491)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3889..3900)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4009..4025)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4010..4026)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    4055..4176
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4066..4085)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4120..4139
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4384..4401)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4555..5143)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4635..4654)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5314..6174)
                     /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     5937..5956
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6175..6279)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6305..6760)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6442..6463)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6654..6673
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc
       61 tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc
      121 actaggggtt ccttgtagtt aatgattaac ccgccatgct acttatctac gtagccatgc
      181 tctaggaaga tccagggccc tgcgtatgag tgcaagtggg ttttaggacc aggatgaggc
      241 ggggtggggg tgcctacctg acgaccggcc ccgacccact ggacaagcac ccaaccccca
      301 ttccccaaat tgcgcatccc ctatcagaga gggggagggg aaacaggatg cggcgaggcg
      361 cgtgcgcact gccagcttca gcaccgcgga cagtgccttc gcccccgcct ggcggcgcgc
      421 gccaccgccg cctcagcact gaaggcgcgc tgacgtcact cgccggtccc ccgcaaactc
      481 cccttcccgg ccaccttggt cgcgtccgcg ccgccgccgg cccagccgga ccgcaccacg
      541 cgaggcgcga gatagggggg cacgggcgcg accatctgcg ctgcggcgcc ggcgactcag
      601 cgctgcctca gtctgcggtg ggcagcggag gagtcgtgtc gtgcctgaga gcgcagtcca
      661 attcaagctg ctagcaagga tcccgccacc atgaaactgg aaatttttag ctggtgggcc
      721 ggcgatgaag gcccggccct ggaagccctg attcgcctgt ataaacagaa atatccgggc
      781 gtggaggtga ttaacgccac cgtgaccggc ggcgccggcg tgaacgcccg cgccgtgctg
      841 aaaacccgca tgctgggcgg cgatccgccg gatacctttc aggtggcagc cggcatggaa
      901 ctgattggca cctgggtggt ggccaaccgc atggaagacc tgagcgccct gtttcgccag
      961 gaaggctggc tgcaagcctt tccgaaaggc ctgattgatc tgattagcta taaaggcggc
     1021 atttggagcg tgccggtgaa cattcatcgc agcaacgtga tgtggtatct gccggccaaa
     1081 ctgaaagaat ggggcgtgaa cccgccgcgc acctgggatg aatttctggc cacctgccag
     1141 accctgaaac agaaaggcct ggaagccccg ctggccctgg gcgaaaactg gacccagcag
     1201 catctgtggg aaagcgtggc cctggctgtg ctgggcccgg atgattggaa caacctgtgg
     1261 aacggcaaac tgaaatttac cgatccgaaa gccgtgcgcg cctgggaagt gtttggccgc
     1321 gtgctggatt gcgccaacaa agatgccgcc ggcctgagct ggcagcaggc cgtggatcgc
     1381 gtggtgcagg gcaaagccgc ctttaacgtg atgggcgatt gggccgccgg ctatatgacc
     1441 accaccctga aactgaaacc gggcaccgat tttgcctggg ccccgagccc gggcacccag
     1501 ggcgtgttta tgatggtgag cgatagcttt ggcctgccga aaggcgccaa aaaccgccag
     1561 aacgccatta actggctgcg cctggtgggc agcaaagaag gccaggatac ctttaacccg
     1621 ctgaaaggca gcattgccgc ccgcctggat agcgatccga gcaaatatcc cgccagccac
     1681 aacgtctata tcatggccga caagcagagg aacggcatca aggcgaactt caagatccgc
     1741 cacaacatcg aggacggcgg cgtgcagctc gcctaccact accagcagaa cacccccatc
     1801 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc caaactttcg
     1861 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     1921 atcactctcg gcatggacga gctgtacaag ggcggtaccg gagggagcat ggtgagcaag
     1981 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     2041 ggccacaagt tcagcgtgtc cggcgagggt gagggcgatg ccacctacgg caagctgacc
     2101 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     2161 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
     2221 ttcaagtccg ccatgcccga aggctacatc caggagcgca ccatcttctt caaggacgac
     2281 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     2341 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcataa gctggagtac
     2401 aactttaaca accccaacgc ctatggccag agcgccatgc gcgattggcg cagcaaccgc
     2461 attgtgggca gcctggtggc aggcgccgtg gccccggaaa gctttatgag ccagtttggc
     2521 accgtgatgg aaatttttct gcaaacccgc aacccgcagg ccgccgccaa cgccgcccag
     2581 gccattgccg atcaggtggg cctgggccgc ctgggccagc tgcaggtcga cgaacaaaaa
     2641 ctcatctcag aagaggatct gaatgctgtg ggccaggaca cgcaggagcg tatggtgtct
     2701 aagggcgaag agctgatcaa ggaaaatatg cgtatgaagg tggtcatgga aggttcggtc
     2761 aacggccacc aattcaaatg cacaggtgaa ggagaaggca atccgtacat gggaactcaa
     2821 accatgagga tcaaagtcat cgagggagga cccctgccat ttgcctttga cattcttgcc
     2881 acgtcgttca tgtatggcag ccgtactttt atcaagtacc cgaaaggcat tcctgatttc
     2941 tttaaacagt cctttcctga gggttttact tgggaaagag ttacgagata cgaagatggt
     3001 ggagtcgtca ccgtcatgca ggacaccagc cttgaggatg gctgtctcgt ttaccacgtc
     3061 caagtcagag gggtaaactt tccttccaat ggtcccgtga tgcagaagaa gaccaagggt
     3121 tgggagccta atacagagat gatgtatcca gcagatggtg gtctgagggg atacactcat
     3181 atggcactga aagttgatgg tggtggccat ctgtcttgct ctttcgtaac aacttacagg
     3241 tcaaaaaaga ccgtcgggaa catcaagatg cccggtatcc atgccgttga tcaccgcctg
     3301 gaaaggttag aggaaagtga caatgaaatg ttcgtagtac aacgcgaaca cgcagttgcc
     3361 aagttcgccg ggcttggtgg tgggatggac gagctgtaca agtagaagct tatcgataat
     3421 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     3481 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     3541 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     3601 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     3661 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     3721 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     3781 ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc
     3841 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     3901 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     3961 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg ataccgtcga
     4021 cctcgacccg ggcggccgct tcgagcagac atgataagat acattgatga gtttggacaa
     4081 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
     4141 ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt
     4201 atgtttcagg ttcaggggga gatgtgggag gttttttaaa gcaagtaaaa cctctacaaa
     4261 tgtggtaaaa tcgataagga tcttcctaga gcatggctac gtagataagt agcatggcgg
     4321 gttaatcatt aactacaagg aacccctagt gatggagttg gccactccct ctctgcgcgc
     4381 tcgctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     4441 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     4501 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     4561 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     4621 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     4681 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     4741 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     4801 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     4861 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     4921 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     4981 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     5041 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     5101 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     5161 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     5221 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     5281 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     5341 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     5401 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     5461 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     5521 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     5581 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     5641 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     5701 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     5761 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     5821 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     5881 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     5941 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     6001 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     6061 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     6121 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     6181 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     6241 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     6301 acctaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     6361 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     6421 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     6481 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     6541 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     6601 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     6661 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     6721 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc attcaggctg
     6781 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgc
//
