LOCUS       Exported                4730 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Fluorescent reporter for membrane voltage with improved brightness.
ACCESSION   .
VERSION     .
KEYWORDS    pBAD_His B_QuasAr1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4730)
  AUTHORS   Hochbaum DR, Zhao Y, Farhi SL, Klapoetke N, Werley CA, Kapoor V, Zou
            P, Kralj JM, Maclaurin D, Smedemark-Margulies N, Saulnier JL, 
            Boulting GL, Straub C, Cho YK, Melkonian M, Wong GK, Harrison DJ, 
            Murthy VN, Sabatini BL, Boyden ES, Campbell RE, Cohen AE
  TITLE     All-optical electrophysiology in mammalian neurons using engineered 
            microbial rhodopsins.
  JOURNAL   Nat Methods. 2014 Aug;11(8):825-33. doi: 10.1038/nmeth.3000. Epub 
            2014 Jun 22.
  PUBMED    24952910
REFERENCE   2  (bases 1 to 4730)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4730
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..285
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     208..227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             331..1089
                     /codon_start=1
                     /product="achaerhodopsin-3 (Arch) derivative with improved 
                     brightness (Hochbaum et al., 2014)"
                     /label=QuasAr1
                     /translation="MDPIALQAGYDLLGDGRPETLWLGIGTLLMLIGTFYFLVRGWGVT
                     DKDAREYYAVTILVSGIASAAYLSMFFGIGLTEVSVGGEMLDIYYARYAHWLFTTPLLL
                     LHLALLAKVDRVTIGTLVGVDALMIVTGLIGALSHTAIARYSWWLFSTICMIVVLYVLA
                     TSLRSAAKERGPEVASTFNTLTALVLVLWTAYPILWIIGTEGAGVVGLGIETLLFMVLD
                     VTAKVGFGFILLRSRAILGDTEAPEPSAGAD"
     primer_bind     complement(1131..1148)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1301..1387
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1479..1506
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1525..1616
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1617..2477
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1835..1854)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2648..3236
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3137..3156
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3390..3407
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3422..3562
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3488..3507)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3648..3670
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3826..4704)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct
       61 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
      121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
      181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg
      241 atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg ttttttgggc
      301 taacaggagg aattaaccat gggttctaga atggacccca tcgctctgca ggctggttac
      361 gacctactgg gtgacggcag acctgaaacc ctgtggctgg gcatcggcac tctgctgatg
      421 ctgattggaa ccttctactt tctggtccgc ggatggggag tcaccgataa ggatgcccgg
      481 gaatattacg ctgtgactat cctggtgtcc ggaatcgcat ccgccgcata tctgtctatg
      541 ttctttggta tcgggcttac tgaggtgtcc gtcgggggcg aaatgttgga tatctattat
      601 gccaggtacg cccattggct gtttaccacc ccacttctgc tgctgcacct ggcccttctc
      661 gctaaggtgg atcgggtgac catcggcacc ctggtgggtg tggacgccct gatgatcgtc
      721 actggcctca tcggagcctt gagccacacg gccatagcca gatacagttg gtggttgttc
      781 tctacaattt gcatgatagt ggtgctctat gttctggcta catccctgcg atctgctgca
      841 aaggagcggg gccccgaggt ggcatctacc tttaacaccc tgacagctct ggtcttggtg
      901 ctgtggaccg cttaccctat cctgtggatc ataggcactg agggcgctgg cgtggtgggc
      961 ctgggcatcg aaactctgct gtttatggtg ttggacgtga ctgccaaggt cggctttggc
     1021 tttatcctgt tgagatcccg ggctattctg ggcgacaccg aggcaccaga acccagtgcc
     1081 ggtgccgatt agaagcttgg ctgttttggc ggatgagaga agattttcag cctgatacag
     1141 attaaatcag aacgcagaag cggtctgata aaacagaatt tgcctggcgg cagtagcgcg
     1201 gtggtcccac ctgaccccat gccgaactca gaagtgaaac gccgtagcgc cgatggtagt
     1261 gtggggtctc cccatgcgag agtagggaac tgccaggcat caaataaaac gaaaggctca
     1321 gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag
     1381 gacaaatccg ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc
     1441 aggacgcccg ccataaactg ccaggcatca aattaagcag aaggccatcc tgacggatgg
     1501 cctttttgcg tttctacaaa ctcttttgtt tatttttcta aatacattca aatatgtatc
     1561 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     1621 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     1681 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     1741 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     1801 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg
     1861 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     1921 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     1981 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     2041 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     2101 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     2161 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     2221 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     2281 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     2341 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     2401 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     2461 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     2521 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     2581 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     2641 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     2701 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     2761 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     2821 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     2881 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     2941 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     3001 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     3061 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     3121 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     3181 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     3241 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     3301 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     3361 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     3421 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca
     3481 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct
     3541 acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg
     3601 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     3661 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagcagatca attcgcgcgc
     3721 gaaggcgaag cggcatgcat aatgtgcctg tcaaatggac gaagcaggga ttctgcaaac
     3781 cctatgctac tccgtcaagc cgtcaattgt ctgattcgtt accaattatg acaacttgac
     3841 ggctacatca ttcacttttt cttcacaacc ggcacggaac tcgctcgggc tggccccggt
     3901 gcatttttta aatacccgcg agaaatagag ttgatcgtca aaaccaacat tgcgaccgac
     3961 ggtggcgata ggcatccggg tggtgctcaa aagcagcttc gcctggctga tacgttggtc
     4021 ctcgcgccag cttaagacgc taatccctaa ctgctggcgg aaaagatgtg acagacgcga
     4081 cggcgacaag caaacatgct gtgcgacgct ggcgatatca aaattgctgt ctgccaggtg
     4141 atcgctgatg tactgacaag cctcgcgtac ccgattatcc atcggtggat ggagcgactc
     4201 gttaatcgct tccatgcgcc gcagtaacaa ttgctcaagc agatttatcg ccagcagctc
     4261 cgaatagcgc ccttcccctt gcccggcgtt aatgatttgc ccaaacaggt cgctgaaatg
     4321 cggctggtgc gcttcatccg ggcgaaagaa ccccgtattg gcaaatattg acggccagtt
     4381 aagccattca tgccagtagg cgcgcggacg aaagtaaacc cactggtgat accattcgcg
     4441 agcctccgga tgacgaccgt agtgatgaat ctctcctggc gggaacagca aaatatcacc
     4501 cggtcggcaa acaaattctc gtccctgatt tttcaccacc ccctgaccgc gaatggtgag
     4561 attgagaata taacctttca ttcccagcgg tcggtcgata aaaaaatcga gataaccgtt
     4621 ggcctcaatc ggcgttaaac ccgccaccag atgggcatta aacgagtatc ccggcagcag
     4681 gggatcattt tgcgcttcag ccatactttt catactcccg ccattcagag
//
