LOCUS       Exported                6274 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  expresses cytoplasmic ATP sensor in astrocytes with red reference 
            protein.
ACCESSION   .
VERSION     .
KEYWORDS    GfaABC1D-cyto-Ruby3-iATPSnFR1.0
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6274)
  AUTHORS   Lobas MA, Tao R, Nagai J, Kronschlager MT, Borden PM, Marvin JS, 
            Looger LL, Khakh BS
  TITLE     A genetically encoded single-wavelength sensor for imaging cytosolic
            and cell surface ATP.
  JOURNAL   Nat Commun. 2019 Feb 12;10(1):711. doi: 10.1038/s41467-019-08441-5.
  PUBMED    30755613
REFERENCE   2  (bases 1 to 6274)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6274
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     1260..1281
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1384..1405)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     polyA_signal    1884..2005
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1895..1914)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1949..1968
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   2167..2296
                     /label=AAV2 ITR
     primer_bind     complement(2315..2332)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2315..2331)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2324..2346)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2473..2928
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2560..2579)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2770..2791
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2954..3058
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3059..3919
                     /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(3277..3296)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4090..4678
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4579..4598
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4832..4849
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4966..4987
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5002..5032
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5040..5056
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     5045..5067
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5064..5080
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5064..5080
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   5118..5247
                     /label=AAV2 ITR
     intron          6098..6230
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     6213..6232
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
ORIGIN
        1 taatacgact cactataggc tagcatggtg tctaagggcg aagagctgat caaggaaaat
       61 atgcgtatga aggtggtcat ggaaggttcg gtcaacggcc accaattcaa atgcacaggt
      121 gaaggagaag gcaggccgta cgagggagtg caaaccatga ggatcaaagt catcgaggga
      181 ggacccctgc catttgcctt tgacattctt gccacgtcgt tcatgtatgg cagccgtact
      241 tttatcaagt acccggcaga cattcctgat ttctttaaac agtcctttcc tgagggtttt
      301 acttgggaaa gagttacgag atacgaagat ggtggagtcg tcaccgtcac gcaggacacc
      361 agccttgagg atggcgaact cgtttacaac gtcaaagtca gaggggtaaa ctttccctcc
      421 aatggtcccg tgatgcagaa gaagaccaag ggttgggagc ctaatacaga gatgatgtat
      481 ccagcagatg gtggtctgag gggatacact gatattgcac tgaaagttga tggtggtggc
      541 catctgcatt gcaatttcgt aacaacttac aggtcaaaaa agaccgtcgg gaacatcaag
      601 atgcccggtg tccatgccgt tgatcaccgc ctggaaagga tagaggaaag tgacaatgaa
      661 acgtacgtag tacaacgcga agtcgcagtt gccaagtact ccaatcttgg tggtgctagc
      721 atgaagacta ttcacgtgag tgtcgtaact cccgacgggc ctgtatatga ggatgacgtt
      781 gaaatggtga gcgtcaaagc aaaaagtggc gagctcggta ttctcccagg ccacattccc
      841 ttgaaagctc ccctagagat cagtgccgca cgcctgaaga aagggggcaa aacacagtat
      901 atcgctgtgt caggcggcaa cctcgaagtg cggcctgaca aggtgaccat ctatgcccag
      961 gcagccgaga gggctgagga tatcgatgtc ctgcgcgcca aggccgccaa ggagagagcc
     1021 gagcgccgac tccaatcaca ggtcctgagc cacaacgtct atatcaccgc cgacaagcag
     1081 aagaacggca tcaaggcgaa cttcaagatc cgccacaacg tggaggacgg cagcatgcag
     1141 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     1201 aaccactacc tgagcaccca gtccgtgctg agcaaagacc ctaacgagaa gcgcgatcac
     1261 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     1321 aagggcggta ccggagggag catgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     1381 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgcg cggcgagggc
     1441 gagggcgatg ccaccaacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     1501 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc
     1561 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     1621 caggagcgca ccatcagctt caaggacgac ggcacctaca agacccgcgc cgaggtgaag
     1681 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     1741 ggcaacatcc tggggcacaa gctggagtac aactttgggt tgcacgacat cgattttaag
     1801 cgcgccgagc tcgcccttaa gcgcgcaatg aataggctct cagttgccga aatgaagtag
     1861 gcggccgctt cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag
     1921 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     1981 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     2041 tcagggggag atgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtaaaat
     2101 cgataaggat cttcctagag catggctacg tagataagta gcatggcggg ttaatcatta
     2161 actacaagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     2221 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
     2281 gcgagcgagc gcgcagcctt aattaaccta attcactggc cgtcgtttta caacgtcgtg
     2341 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
     2401 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
     2461 atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     2521 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     2581 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     2641 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     2701 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     2761 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     2821 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     2881 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt
     2941 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     3001 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     3061 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     3121 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     3181 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     3241 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     3301 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     3361 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     3421 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     3481 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     3541 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     3601 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     3661 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     3721 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     3781 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     3841 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     3901 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     3961 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     4021 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     4081 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     4141 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     4201 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     4261 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     4321 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     4381 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     4441 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     4501 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     4561 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     4621 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     4681 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     4741 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     4801 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     4861 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
     4921 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
     4981 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
     5041 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccagatttaa
     5101 ttaaggcctt aattaggctg cgcgctcgct cgctcactga ggccgcccgg gcaaagcccg
     5161 ggcgtcgggc gacctttggt cgcccggcct cagtgagcga gcgagcgcgc agagagggag
     5221 tggccaactc catcactagg ggttccttgt agttaatgat taacccgcca tgctacttat
     5281 ctacgtagcc atgctctagg aagatctaac atatcctggt gtggagtagg ggacgctgct
     5341 ctgacagagg ctcgggggcc tgagctggct ctgtgagctg gggaggaggc agacagccag
     5401 gccttgtctg caagcagacc tggcagcatt gggctggccg ccccccaggg cctcctcttc
     5461 atgcccagtg aatgactcac cttggcacag acacaatgtt cggggtgggc acagtgcctg
     5521 cttcccgccg caccccagcc cccctcaaat gccttccgag aagcccattg agcagggggc
     5581 ttgcattgca ccccagcctg acagcctggc atcttgggat aaaagcagca cagcccccta
     5641 ggggctgccc ttgctgtgtg gcgccaccgg cggtggagaa caaggctcta ttcagcctgt
     5701 gcccaggaaa ggggatcagg ggatgcccag gcatggacag tgggtggcag ggggggagag
     5761 gagggctgtc tgcttcccag aagtccaagg acacaaatgg gtgaggggag agctctcccc
     5821 atagctgggc tgcggcccaa ccccaccccc tcaggctatg ccagggggtg ttgccagggg
     5881 cacccgggca tcgccagtct agcccactcc ttcataaagc cctcgcatcc caggagcgag
     5941 cagagccaga gcaggttgga gaggagacgc atcacctccg ctgctcgcaa gctttattgc
     6001 ggtagtttat cacagttaaa ttgctaacgc agtcagtgct tctgacacaa cagtctcgaa
     6061 cttaagctgc agaagttggt cgtgaggcac tgggcaggta agtatcaagg ttacaagaca
     6121 ggtttaagga gaccaataga aactgggctt gtcgagacag agaagactct tgcgtttctg
     6181 ataggcacct attggtctta ctgacatcca ctttgccttt ctctccacag gtgtccactc
     6241 ccagttcaat tacagctctt aaggctagag tact
//
