LOCUS       Exported                6778 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Astrocyte expression of fluorescent sensor for extracellular 
            lactate.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-GFAP-eLACCO1.1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6778)
  TITLE     A genetically encoded fluorescent biosensor for extracellular 
            L-lactate
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6778)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6778
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   651..780
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     primer_bind     2409..2430
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2536..2557)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2797..2816)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    3555..4143
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3608..3628)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4026..4037)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4175..4651
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4374..4393)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4691..4831
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4691..4820
                     /label=AAV2 ITR
     rep_origin      4906..5361
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4993..5012)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5203..5224
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5378..5397)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5497..5519
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5557..5575)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5643..5747
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5748..6608
                     /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(5966..5985)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt cctgcaggca
      661 gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt ggtcgcccgg
      721 cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct
      781 gcggccgcac gcagatctaa catatcctgg tgtggagtag gggacgctgc tctgacagag
      841 gctcgggggc ctgagctggc tctgtgagct ggggaggagg cagacagcca ggccttgtct
      901 gcaagcagac ctggcagcat tgggctggcc gccccccagg gcctcctctt catgcccagt
      961 gaatgactca ccttggcaca gacacaatgt tcggggtggg cacagtgcct gcttcccgcc
     1021 gcaccccagc ccccctcaaa tgccttccga gaagcccatt gagcaggggg cttgcattgc
     1081 accccagcct gacagcctgg catcttggga taaaagcagc acagccccct aggggctgcc
     1141 cttgctgtgt ggcgccaccg gcggtggaga acaaggctct attcagcctg tgcccaggaa
     1201 aggggatcag gggatgccca ggcatggaca gtgggtggca gggggggaga ggagggctgt
     1261 ctgcttccca gaagtccaag gacacaaatg ggtgagggga gagctctccc catagctggg
     1321 ctgcggccca accccacccc ctcaggctat gccagggggt gttgccaggg gcacccgggc
     1381 atcgccagtc tagcccactc cttcataaag ccctcgcatc ccaggagcga gcagagccag
     1441 agcaggttgg agaggagacg catcacctcc gctgctcgcg gggatccacc ggtgagctcc
     1501 cgcggcggcc gctctagatt agtctgcaga atcaccatgg gaatccaagg agggtctgtc
     1561 ctgttcgggc tgctgctcgt cctggctgtc ttctgccatt caggtcatag cagatctatg
     1621 ttctctccct tggcagtggc ccaggctcga cgctataggt ggaggattca aaccgcgtgg
     1681 gatgctggaa cagtaggcta cagtctgttc caaaagttta cggagcgggt aaaggaactg
     1741 acggatggac agcttgaggt ccagcctttt ccagcgggtg cagtcgtagg cacgtttgat
     1801 atgttcgacg cggtcaagac tggagttttg gacggaatga atcctttcac gctgttttgg
     1861 gcagggagga tgccggttac agctttcctc tccagctacg ccctcgggtt ggaccgacct
     1921 gaccagtggg aaacatggtt ttactccctg ggtggcctcg acaacgcccg acgagccttt
     1981 gccgagcaag gcctcttcta cgtgggacca gttcagcacg acctcaatac aattcattca
     2041 aggaagccga tccggagatt tgaggacttc aaaggtgtca agctccgagt gccgggcggc
     2101 atgattgcgg aagtttttgc ggccgcagga gcttccaccg tcctcttgcc tgggggggag
     2161 gtctaccccg ctctggaaag aggggtaata gattggagcc acaacgtcta tatcatggcc
     2221 gacaagcaga ggaacggcat caaggcgaac ttcgagatcc gccacaacat cgaggacggc
     2281 ggcgtgcagc tcgcctacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     2341 ctgcccgaca accactacct gagcacccag accaaacttt cgaaagaccc caacgagaag
     2401 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     2461 gagctgtaca agggcggtac cggagggaga atggtgagca agggcgagga gctgttcacc
     2521 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     2581 tccggcgagg gtgagggcga tgccacctcc ggcaagctga ccctgaagtt catctgcacc
     2641 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag
     2701 tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc ggccatgccc
     2761 gaaggctaca tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     2821 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     2881 ttcaaggagg acggcaacat cctggggcat aagctggagt acagctttaa cgatggtggt
     2941 gcggctgatt ttgttgggcc tgcagtaaac tataacttgg ggttccatca ggtggcaaag
     3001 tacatcataa tggggcctcc agagacgcct gccattcatc aacctgtaga cctcatggac
     3061 ttcaccataa accttaatcg gtggcgcagc cttccaaagc ctctgcaaga aagattcatc
     3121 gcggcggttc acgagtactc ttggattcac tacgcaggca ttcagaaggc taatttggag
     3181 gcgtggccga aatatcggca ggcaggtgtt gaggtcatcc ggctgagcaa cgaggatgtt
     3241 agaaaattca ggcgactcgc tattcccatt tggtttaagt gggctaagat ggataaatac
     3301 tcacgagagg cattcgcgtc ccaactggag tatatgaaag gtataggtta cgttacggac
     3361 gaggaattga agggcttgag cctgggctcc acctccggct ccggcaagcc cggctccggc
     3421 gagggctcca ccaagggcga attccttgaa aatggtggga catccttatc agagaaaaca
     3481 gttcttctgc tggtgactcc atttctggca gcagcctgga gccttcatcc ctaaggatca
     3541 tcaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     3601 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     3661 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     3721 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     3781 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     3841 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     3901 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct
     3961 ttccttggct gctcgcctat gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     4021 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
     4081 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
     4141 cgcatcgata ccgagcgctg ctcgagagat ctacgggtgg catccctgtg acccctcccc
     4201 agtgcctctc ctggccctgg aagttgccac tccagtgccc accagccttg tcctaataaa
     4261 attaagttgc atcattttgt ctgactaggt gtccttctat aatattatgg ggtggagggg
     4321 ggtggtatgg agcaaggggc aagttgggaa gacaacctgt agggcctgcg gggtctattg
     4381 ggaaccaagc tggagtgcag tggcacaatc ttggctcact gcaatctccg cctcctgggt
     4441 tcaagcgatt ctcctgcctc agcctcccga gttgttggga ttccaggcat gcatgaccag
     4501 gctcagctaa tttttgtttt tttggtagag acggggtttc accatattgg ccaggctggt
     4561 ctccaactcc taatctcagg tgatctaccc accttggcct cccaaattgc tgggattaca
     4621 ggcgtgaacc actgctccct tccctgtcct tctgattttg taggtaacca cgtgcggacc
     4681 gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg
     4741 ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca
     4801 gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg
     4861 catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg
     4921 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     4981 ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     5041 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     5101 tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga
     5161 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     5221 ctggaacaac actcaactct atctcgggct attcttttga tttataaggg attttgccga
     5281 tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     5341 aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat
     5401 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     5461 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     5521 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
     5581 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     5641 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     5701 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     5761 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     5821 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     5881 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     5941 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     6001 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     6061 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     6121 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     6181 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     6241 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     6301 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     6361 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     6421 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     6481 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     6541 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     6601 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     6661 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     6721 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttc
//
